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The retinol-binding protein from porcine serum has been isolated and characterized. Its molecular weight and amino-acid composition were determined and compared to these data from its human, monkey and rat counterpart. In anology to these proteins pig retinol-binding protein is under physiological conditions bound to a larger plasma protein and has a carboxyl-terminal residue of arginine suggesting a general mechanism for the delivery of vitamin A to the tissue cells.

A colony counting method was used to record the rate of growth of the T1 strain ofMycoplasma mycoides in various media. A maximum population of 5 × 109 C.F.U./ml. was consistently obtained in tryptose serum broth, and a population of 5 × 108 C.F.U./ml. was supported for at least 3–4 days. The substitution of bovine for porcine serum in this medium made no difference to the growth of the organism. Several media appeared to support 105–106 C.F.U./ml. for an incubation period of 8 weeks. C.F.U.=Colony forming unit.

Summary and Conclusions1. Some of the nutritional requirements for the in vitro cultivation of hog kidney epithelial cells were determined. The data indicate that cultures grown in a “simple” medium containing Synthetic Mixture #199 (92.5%) and hog serum (7.5%) were consistently more proliferative than cultures cultivated in more complex media containing embryo extract, serum ultra-filtrate and serum of homologous and heterologous origin. 2. None of the media tested adequately supported growth of either splenic or testicular fibroblasts. 3. The influence of various Porcine Serum Fractions (II, IV, V and IV + V) on growth of hog kidney epithelial cells was also investigated. When Fraction IV -f- V was used, excellent growth was achieved. When Fraction II, IV or V was used independently, growth inferior to that of whole serum resulted.

Perry (1964) has shown that piglets failed to absorb homologousBrucella abortusantibodies, whereas they absorbed homologousSalmonella pullorumantibodies much more readily. WhenS. pullorumantibody was used, piglets showed marked differences in their efficiency of absorption. These differences existed whether the antibodies were administered in serum or in colostrum (Perry and Watson, 1967a, b). Moreover, they could not be entirely accounted for by variation in concentration of antibody in the dose, age at dosage, genetical effects or maternal effects (Perry and Watson, 1967a).

Swine with a hemophilia-like disease characterized by a long bleeding time and low antihemophilic factor (AHF, factor VIII) level were infused with porcine plasma and porcine serum. Unlike the response of classical hemophiliacs, but similar to that of patients with von Willebrand's disease, the initial postinfusion levels of AHF were sustained with some fluctuation for 24 hr. Serum infusion resulted in a delayed increase in AHF which reached a maximum concentration between the 12th and 24th hr after infusion. Within 75 hr the effects of both the plasma and serum infusions had vanished. No correction of the prolonged bleeding times was observed following the infusions. The AHF levels encountered after the infusion of plasma and serum cannot be accounted for by the AHF activity contained in the infused materials; new AHF synthesis or AHF release from storage are suggested as possible mechanisms.

A serological survey was carried out to detect hemagglutination-inhibition (HI) antibody against hemagglutinating encephalomyelitis virus (HEV) from the serum of pig. A total of 6, 797 porcine serum samples were collected from 45 prefectures in Japan. As a result, 3, 393 samples (49.9%) had the antibody. Of them, 1, 992 samples (58.7%) had an HI titer of 1:1280 or high. The positive rate varied from 39.6 to 72.0% in each age-group. It was 53.7, 47.0, 39.6, 52.5, 54.4, and 72.0% in the 0-2, 2-4, 4-6, 6-8, 8-l2, and over-12-month groups, respectively. In short, the positive rate became higher in accordance with the advance in age. The occurrence of such a high positive rate as 53.7% in the 0-2 month group, and the same HI titers of suckling piglets as those of their mother sows may be due to the absorption of antibodies from colostrum. The breed of pigs may not be a factor to induce HEV infection. The positive rate was high in the Kinki and Chugoku districts (66.1 and 65.1%, respectively) and low in the Tohoku and Kanto districts (26.6 and 27.8%, respectively). Antibodies against HEV were proved in Japan in 1965 and confirmed in every subsequent year. In 1973, more than 60.0% of the pigs examined were found to possess those antibodies. Therefore, it is concluded that HEV infection may be widespread in Japan.

Summary The development of indirect haemagglutinins against M. hyopneumoniae in serum and tracheobronchial secretions was investigated in experimentally inoculated pigs. IHA-antibodies against M. hyopneumoniae were demonstrated in serum 2–4 weeks p. i. and reached maximum levels about 8–9 weeks p. i. Immunochemical characterization of the serum antibody response in one pig showed that the IHA-antibodies were both IgG and high molecular IgA. IHA-antibodies were present in tracheobronchial secretions of all pigs investigated 2 weeks p. i. These pigs and 2 pigs investigated 4 weeks p. i. had IHA-titres in the tracheobronchial secretion without having detectable amounts of antibodies in serum. Immunochemical characterization of this early antibody response in one pig showed that the IHA-antibodies were high molecular IgA probably secretory IgA. Later in the experiment the IHA-titres in tracheobronchial secretions were as high as or higher than those found in serum. When the experiment was finished at 13 weeks p. i. the IHA-antibodies in tracheobronchial secretions of one pig investigated were predominantly high molecular IgA. Zusammenfassung Immunreaktionen im Serum und Tracheobronchialsekret von experimentell mit Mycoplasma hyopneumonia infizierten Schweinen An experimentell infizierten Schweinen ist die Entwicklung von Antikorpertitern gegen M. hyopneumonias untersucht worden. Zum Nachweis der Antikorperwerte wurde der indirekte Hamagglutinationstest (IHA) verwendet. 2–4 Wochen p. inf. konnten Antikorper bereits im Serum nachgewiesen werden. Die Serumtiter erreichten in der 8. und 9. Woche ihren Hohepunkt. Die immunochemische Charakterisierung der Serumantikorper eines Schweines zeigte, das die IHA-Titer von IgG und hoch-molekularen IgA gebildet wurden. Im Tracheobronchialsekret liesen sich IHA-Antikorper bei allen Schweinen ab der 2. Woche p. inf. nachweisen. Diese Tiere, sowie 2 Schweine, die in der 4. Woche p. inf. untersucht worden waren, besasen noch keine Serumantikorper. Immunochemisch wurde festgestellt, das diese fruhen Antikorper im Tracheobronchialsekret ausschlieslich hochmolekulare, wahrscheinlich sekretorische IgA waren. Im weiteren Verlauf der Versuche waren die IHA-Titer im Tracheobronchialsekret entweder genauso hoch oder hoher als im Serum. Schlieslich ist in der 13. Woche p. inf. nochmals das Tracheobronchialsekret eines Schweines untersucht worden. Auch dieses Sekret enthielt vorwiegend hochmolekulare IgA. Resume Reactions immunitaires dans le serum et la secretion tracheobronchiale chez des porcs infectes experimentalement avec Mycoplasma hyopneumoniae On a examine le titre des anticorps contre M. hyopneumoniae chez des porcs infectes experimentalement. On a utilise le test d'inhibition de l'hemagglutination (IHA) pour la recherche des anticorps. On a pu montrer la presence d'anticorps dans le serum deja 2–4 semaines apres l'infection. Les titres serologiques ont atteint leur maximum dans la 8eme et 9eme semaine. La caracterisation immunochimique des anticorps seriques d'un porc a montre que le titre d'inhibition de l'hemagglutination dependait des Ig G et des IG A a poids moleculaire eleve. Les anticorps inhibant l'hemagglutination ont ete trouves dans la secretion tracheobronchiale chez tous les porcs des la 2eme semaine apres l'infection. Ces animaux et 2 porcs, examines 4 semaines apres l'infection, ne possedaient encore aucun anticorps serique. L'analyse immunochimique a etabli que ces anticorps precoces des secretions tracheobronchiales etaient exclusivement d'un poids moleculaire eleve et vraisemblablement des Ig A de secretion. Les titres des anticorps inhibang l'hemagglutination des secretions tracheobronchiales furent au cours des recherches soit aussi elves soit plus eleves que ceux des anticorps seriques. On a enfin encore examine la secretion tracheobronchiale d'un porc dans la 13eme semaine apres l'infection: cette secretion contenait avant tout des Ig A de poids moleculaire eleve. Resumen Reaccion inmunologica en el suero sanguineo y secrecion traqueobronquial de cerdos infectados experimentalmente con Mycoplasma hyopneumoniae En cerdos infectados experimentalmente se estudio el desarrollo de los titulos de anticuerpos frente a M. hyopneumoniae. Para la deteccion de los valores de anticuerpos se utilizo la prueba de hemoaglutinacion indirecta (HAI). De 2–4 semanas p. inf. ya se pudieron identificar anticuerpos en el suero sanguineo. Los titulos sericos alcanzaron su fastigio durante la 8° y 9° semana. La caracterizacion inmunoquimica de los anticuerpos sericos de un cerdo revelo que los titulos HAI eran formados por IgG e IgA polimolecular. En la secrecion traqueobronquial se pudieron identificar anticuerpos HAI en todos los cerdos a partir de la 2° semana p. inf. Estos animales, asi como 2 cerdos que se examinaron en la 4° semana p. inf., aun no poseian anticuerpos sericos. Se comprobo inmunoquimicamente que estos anticuerpos precoces en la secrecion traqueobronquial eran exclusivamente IgA polimolecular, tal vez secretorio. En el curso subsiguiente de los ensayos eran los titulos HAI en la secrecion traqueobronquial bien igual o mas elevados que en el suero sanguineo. Por ultimo, se analizo de nuevo en la semana 13° p. inf. la secrecion traqueobronquial de un cerdo. Tambien esta secrecion contenia sobre todo IgA polimolecular.

Summary Leukocyte-specific antigens were demonstrated using antisera induced in rabbits by the intradermal injections of a thermostable, ethanol-insoluble fraction of porcine leukocytes, mixed with Freund adjuvants. Positive results were obtained in complement fixation, tanned cell hemagglutination and double diffusion gel precipitation tests. These antigens were absent from similar fractions of porcine serum, thrombocytes, erythrocytes, spleen, kidney, liver, muscle, brain, testicle, and adrenal. In gel precipitation tests, two leukocyte-specific lines were observed; in immunoelectrophoresis, both of them were located in the anodal part of the electrophoretic field. Cross reactions were observed with a thermostable fraction of bovine leukocytes. Negative leukoagglutination tests indicated that the thermostable, leukocyte-specific antigens appear inside the leukocytes rather than on their surface. Resume Des antigenes leucocytaires specifiques ont ete mis en evidence au moyen d'antiserums de lapins, prepares par injection intra-dermique de fractions de leucocytes porcins, thermostables et insolubles dans l'alcool ethylique, melanges avec de l'adjuvant de Freund. Des resultats positifs ont ete obtenus au moyen des tests de fixation du complement, de l'hemagglutination passive et de la double diffusion passive en gelose. L'etude de fractions semblables obtenues a partir de tissus de porc: soit du serum, des thrombocytes, des erythrocytes, de la rate, du rein, du foie, du muscle, du cerveau, du testicule et de la surrenale n'a pas demontre ces antigenes. Dans les tests de precipitation en gelose, deux lignes specifiques pour les leucocytes ont ete decelees: a l'analyse immuno-electrophoretique, ces deux lignes ont ete localisees a la partie anodique du champs electrophoretique. Des reactions croisees ont ete observees avec une fraction thermostable des leucocytes bovins. L'absence de leuco-agglutination permet de conclure que ces antigenes leucocytaires specifiques, se trouvent a l'interieur plutot que sur la surface de ces cellules. Zusammenfassung Kaninchen wurden durch intradermal Injektion einer thermostabilen, nicht alkoholloslichen Fraktion von Schweineleukozyten gemischt mit Freundschem Adjuvans immunisiert. Mit diesen Antiseren gelang es leukozytenspezifische Antigene nachzuweisen. Positive Ergebnisse wurden in der Komplementbindungsreaktion der Hamagglutination tannierter Erythrozyten und in der Doppeldiffusionsmethode der Immunprazipitationsreaktion erzielt. Diese Antigene konnten jedoch nicht in analogen Fraktionen von Schweineserum, Thrombozyten, Erythrozyten, Milz, Niere, Leber, Muskel, Hirn, Hoden und Nebenniere nachgewiesen werden. In der Prazipitationsreaktion im Agargel waren zwei leukozytenspezifische Linien zu erkennen, die in der Immunelektrophorese im anodischen Wanderungsbereich zur Darstellung kamen. Mit einer thermostabilen Fraktion von Rinderleukozyten waren Kreuzreaktionen zu beobachten. Der negative Ausfall der Leukozytenagglutinationsreaktion deutet darauf hin, das die thermostabilen, leukozytenspezifischen Antigene im Zellinnern und nicht an der Zelloberflache lokalisiert sind.

PORCINE serum is particularly rich in macroglobulins: we have found it to contain 8 per cent of components with sedimentation coefficients up to approximately 17 S (Fig. 1). Gel filtration with ‘Sephadex G200’ offers a useful means for the preparation of macroglobulins1–3 and preparative zone electrophoresis allows a good separation of the γ1M and α2M globulins.

The isolation and immunologic characterisation of a porcine serum immunoglobulin is described. The isolated protein is shown to be distinct from IgG1, IgG2 and IgM but to share antigenic determinants with a colostral immunoglobulin, thought to be equivalent to human secretory IgA, which possesses additional determinants. It is suggested that the isolated serum immunoglobulin is equivalent to human serum IgA and that the pig possesses an IgA system analogous to that found in man.

The physicochemical properties of natural phage-neutralizing antibodies were studied. Natural neutralizing antibodies against phages ΦX 174 and T 2 were found in the 19 S fraction isolated from normal pig serum by gel filtration on a Bio-Gel P-300 column. The 7 S fraction of normal pig serum possessed no neutralizing activity. The neutralizing activity of the 19 S fraction against phage ΦX 174 was not modified either by inactivation or by the addition of neutralizing cofactor; its activity against phage T 2 was lost by inactivation, but was restored by the addition of cofactor. The neutralizing activity of the 19 S fraction of normal pig serum was completely destroyed by 2-mercaptoethanol and was not restored by the subsequent addition of antibody cofactor. The results of attempts to release phage ΦX 174 from the neutralization complex with normal porcine serum 19 S macroglobulin antibody by the dilution method were the same as those of attempts to dissociate phage from the complex with 7 S type hyperimmune antibody. The virus particle was firmly and irreversibly adsorbed to both types of antibody and was not released by dilution. It is concluded from the results that neutralization of phage ΦX 174 by 19 S macroglobulin molecule of antibody is a simple, irreversible process, for which the thermolabile nonspecific serum components are not required.

An analysis of the immunoglobulin region of porcine blood serum proved the presence of a number of protein fractions which is higher than in other animal species. The presence of antibody activity to tetanus anatoxin examined by radioimmunoelectrophoresis was found in three γ G-globulin fractions, which could be distinguished from each other, and also in two additional fractions. One of the latter was the γ-macroglobulin fraction and the other one might correspond to γ A-globulin even though its mobility was higher than the mobility of γ A-globulin of human serum. A comparison of the immunoglobulin spectrum of porcine serum, colostrum, and of the suckling pig serum after ingestion of the colostrum showed that the main component of porcine colostrum constitutes the γ G-immunoglobulin, even though the colostrum seems to contain other components of the latter which have a higher electrophoretic mobility. The antibody activity of colostrum was found only in fractions γ G and γ M. The fraction of colostrum which might correspond to γ A did not display any activity at all. The activity in suckling pig serum was also concentrated in fractions γ G and γ M.

The very low-density lipoprotein was isolated from porcine serum to examine its chemical and physical properties. Preparations were made by both preparative and zonal ultracentrifuges and the yield was 39 ± 8 mg/100 ml of serum. The lipid content was 92.1 ± 0.7% and the molecular weight was (19.2 ± 1.0)·106. The amino acid content of the apoprotein was similar to that of human very low-density apoprotein. Partial delipidation of the lipoprotein with ethyl ether produced one to three protein-lipid complexes of diminished size. The major complex had a molecular weight of (2.47 ± 0.15) · 105. Fractions isolated from partially delipidated very low-density lipoprotein differed in amino acid composition.

We have demonstrated that sheep red blood cells can be passively sensitized with an extract from Erysipelothrix rhusiopathiae . Diagnostic differential hemagglutination titrations may then be made with porcine serum for Erysipelas antibody.