ALBERT

Notizen: A healthy 22-year-old woman was noted to have erythrocytes of the Pk phenotype: a strong Pk antigen, no detectable P antigen and anti-P antibody in her serum. Her erythrocytes contained four to six times as much Pk glycolipid (globotriaosylceramide or CTH) and approximately half as much P glycolipid (globotertraosylceramide or globoside) as normal red cells. The structures of CTH and globoside were characterized by analysis of permethylated sugars and complement fixation in addition to chromaographic mobility and sugar composition. Inasmuch as the erythrocytes of two Pk individuals that were analysed previously (Marcus et al., 1976) contained no detectable globoside, these abnormalities appear o represent a new phenotype in the P blood group system.

Notizen: We studied 201 unrelated French Basque individuals for HLA and Bf polymorphisms. The haplotypes of eighty-seven of them were deduced from family studies. The results show the frequency of the Bf F1 allele (0.1393) which is the highest one currently reported. They confirm the high frequencies of HLA-Aw19.2 and B18 previously reported in that population and show that a whole haplotype with strong linkage disequilibria, namely Aw19.2, Cw5, B18, Bf F1, DRw3 is frequent. On the other hand, the gene frequency of Bf S is decreased (0.5497) as compared with the other European Caucasoïd populations, while a slight increase in the Bf F gene frequency (0.2960) appears. These results point out that it is of importance to consider the genetic background in choosing the population where linkage disequilibria are to be studied.

Notizen: The genetic control of hybrid resistance to BALB/c fibrosarcoma Meth-A was investigated. A Meth-A tumour grew slower in (BALB/c X C57BL/6)F1 and reciprocal hybrid mice than in syngeneic BALB/c mice and was also found to grow slower in females than in males. Significant F1 resistance was demonstrated after both subcutaneous and intraperitoneal injection of tumour cells. However, (BALB/c X DBA/2)F1 mice did not show any significant resistance to Meth-A. In H-2 linkage studies of [BALB/c X (BALB/c X C57BL/6)] backcross mice, no statistically significant differences in the resistance of H-2 heterozygotes and homozygotes to Meth-A were observed. These results indicated that F1 hybrid resistance to Meth-A was controlled by non-H-2-linked resistance factor(s). No linkage was observed between resistance to Meth-A and coat colour c- and b-loci.

Notizen: CBA/N mice have an X-linked B cell defect which prevents them from responding to non-mitogenic thymic independent (TI-II) antigens such as dinitrophenylated (DNP-AGG) Ficoll. The F1 male progeny of CBA/N female mice express the same defect. Spleen cell suspensions from such defective mice (CBA/N X C3H/HeN F1 males) could not respond to DNP-AGG-Ficoll following in vitro immunization and subsequent transfer into irradiated, syngeneic, F1 male recipients as expected. In contrast, normal CBA/N X C3H/HeN F1 female spleen cells could respond and effect a ‘rescue'; they mounted strong plaque-foriming cell 7 days after in vitro exposure to DNP-AGG-Ficoll and subsequent transfer into irradiated F1 male recipients. Defective F1 male spleen cells could bind significant quantities of DNP-AGG-Ficoll, however, after, in vitro exposure. Extensive washing of these spleen cells could not reverse this binding. Such DNP-AGG-Ficoll-exposed and washed F1 male spleen cells could, after transfer, aid normal untreated F1 female cells in their rescue function. The defective F1 male spleen cells could convey immunogenic quantities of DNP-AGG-Ficoll to the ‘rescuing’ F1 female cells.Mitomycin treatment of F1 male cells did not interfere with their conveyor function. Goat anti-mouse μ serum impeded the passive antigen conveyor function of defective F1 male cells as did prior exposure to high concentrations of free DNP-AGG hapten. Our data support the view that the B cell defect of CBA/N X C3H/HeN F1 male mice does not relate to antigen binding, but rather to an inability to be effectively triggered by certain cell-bound polymeric antigens.

Notizen: Results of a population study with all currently available B-cell specific alloantisera indicate that eight antigens controlled by the RhLA-linked DR locus can now be identified. This leaves a gene frequency of about 0.15 for unidentified or ‘blank’ antigens of that locus. Of the nine identifiable la antigens which are not controlled by the DR locus, three or four may form the basis of a second series which is probably also controlled by the RhLA region.

Notizen: Induction of tolerance to bovine serum albumin was studied in mice selected for high (H) or low (L) antibody responsiveness and in their F1 hybrids. No high or low zone tolerances were obtained in H mice whereas L mice were susceptible to tolerance induction by the two schedules. H mice were immunized by repeated injections of tolerogenic BSA for low zone tolerance induction but not after the administration of a single high dose of tolerogenic BSA. Resistance to tolerance induction is dominant in F1 hybrids.

Notizen: An attempt is made to account for immunoglobulin chain synthesis in terms of genetic events involving IS or controlling elements analogous to those found in bacteria, maize and drosophila. Transposition of variable and constant genes and normal immunoglobulin chain synthesis as well as qualitative and quantitative abnormalities might be explained by such regulatory elements. Intrachromosomal transpositions over short distances would be expressed as apparent hypermutability or redundancy of the variable DNA segment. The constant gene might comprise four sequences coding for the three homology domains and the hinge, separated by intervening sequences. A strong preference for shortrange transposition on the same chromosome and immobilization of the controlling element in the end might account for allelic exclusion.

Notizen: Fab2 fragments from antisera raised in rabbits with partially purified cellular and serum HLA antigens were tested for their ability to block the cytolytic activity of operationally specific HLA-A, B alloantisera. One Fab2 fragment preparation blocked the cytolytic activity of all the HLA-A,B alloantisera tested; the remaining nine inhibited the lytic activity of alloantisera to certain HLA-A,B allospecificities, suggesting that these xenoantisera contain antibody to certain HLA-A,B allotype determinants or to closely associated structures. In contrast to previous reports in the literature none of the xenoantisera contained significant amounts of antibodies to human β2-microglobulin.

Notizen: Immunoglobulin was obtained from a hybridoma cell line, which was a reclone of a hybrid between a rabbit cell and mouse myeloma cell (X63-Ag8). The immunoglobulin, isolated from the cell culture medium was found to be homogeneous by isoelectrofocusing and immunoelectrophoresis and consisted of mouse heavy and rabbit light chains, linked by disulphide bonds. All immunoglobulin molecules carried both mouse and rabbit determinants; mouse determinants were associated only with the heavy chains while rabbit determinants were only associated with the light chains. The rabbit light chains were of Ab4 allotypical specificity, but possess only some of the Ab4 determinants normally present in Ab4/Ab4 animals. It was sugested that the restriction in allotypical specificity may be a general property of light chain Ab allotypes; the normal serum immunoglobulins may be heterogeneous with respect to the allotypic determinants and any one molecule may possess only a proportion of determinants detected by a conventional anti-allotype antiserum.

Notizen: In a family study of coeliac disease, HLA types in fifty-three patients and their relatives were examined. There are no differences in HLA frequencies between child and adult patients. Comparison with a random series of normal controls shows increases frequencies in patients of HLA-A1 and B8, while the family material shows that there is also an excess of haplotype I-8. The excess of homozygotes is thought not to be a factor in the aetiology. Intrafamilial analysis shows that only B8 is significantly associated with the disorder. It is argued that the HLA association does not indicate a ‘coeliac gene’ but that the B8 allele is a major gene in a polygenic system affecting the disorder.

Notizen: Several interesting and clinically important strains of Mus musculus were serotyped with alloantisera recognizing the different alleles of the Thy-1, Ly-1, Ly-2, Ly-3, Ly-4, Ly-5, Ly-6 and Ly-7 loci. The strains were the Biozzi high and low responder strains, of importance in immune responsiveness studies, and the strains BXSB, MRL and MRL—lpr/lpr—strains which spontaneously develop immune complex disease. In addition the related species Mus musculus castaneus was typed with the same reagents. The knowledge of the cell surface phenotype should prove useful in various functional studies involving these interesting strains.

Notizen: Mouse strains differ widely in their natural killer(NK)-cell activity. In the (A X B6) X A backcross, high reactivity was linked to H-2b, although non-H-2-linked genes were also demonstrated (Petranyi et al., 1975). Harmon et al. (1977) demonstrated an H-2Dd-associated reactivity gene (1977). In the present study, we have tested eleven B10 congenic strains for NK activity. The H-2Dd strains B10.A, B10.T(6R), B10.S(7R), B10.HTT and B10.D2 were more highly reactive than B10, B10.S, B10.G, B10.A(2R) and B10.BR, which do not carry the d allele at the H-2D locus. While this confirms the H-2Dd association of a reactivity gene, an exception was found in the B10.A(5R) strain that was low reactive in spite of the fact that it carries H-2Dd. This suggests the possibility that the H-2Dd-associated gene is outside H-2, to the right of Tla.The AKR.H-2b congenic line had the same low activity as the AKR.H-2k strain; both were much lower than B6. This suggests either one of two possibilities: the H-2b-linked reactivity gene is relatively distant from the H-2 complex, although localized on chromosome 17 or alternatively, if localized within or in the close neighbourhood of H-2, it requires non-H-2 genes for full expression.Previously, we have shown that the B6 X DBA/2 F1 hybrid was more highly reactive than either one of its two parental strains (Klein et al., 1978). A similar complementation effect is described in the present paper for the B10.D2 congenic strain. The high reactivity of this line can be due to the combined effect of an H-2b-linked gene from DBA/2 and the non-H-2 background of B10 or, alternatively, the former, together with an H-2b-linked gene from B6 that lies well outside outside the H-2 locus itself.

Notizen: The K region of the H-2 major histocompatibility complex (MHC) of mice of H-2Kk haplotype codes for two distinct alloantigens. One of these alloantigens, designated k-common, is expressed by C3HfB/HeN mice (C3Hf). The other alloantigen, designated k-unique, is not expressed by C3Hf mice. The H-2 haplotype of C3Hf mice has been classified as kv1 and the variant antigen distinguishing this strain from mice of H-2Kk haplotype has been designated kv1-unique. Several transplacentally-induced lung tumours of C3Hf mice express the k-unique, rather than the expected kv1-unique, antigen. The immunogenicity of the k-unique antigen on C3Hf-derived lung tumours varies with different tumours. In particular, the capacity of the k-unique antigen to induce radioresistant immunity in C3Hf mice appears to be lost on long term cultured tumour cells even though the tumour remains susceptible to in vivo immune responses directed against the k-unique antigen. Alterations in expression and in immunogenicity of unique H-2-coded antigens may dictate the nature and efficacy of immune surveillance of autochthanous tumours.

Notizen: The inbred mouse strains BN/a and BN/b have haplotype H-2bp characterized by H-2.33 and lacking any other private specificity known in inbred strains. Two transplantable B cell leukaemias which originated in BN/a and BN/b mice treated with anti-lymphocyte globulin were tested serologicaly for H-2 antigens. Tests during passages 169-181 revealed several quantitatively different reactions with sera against public specificities, some of these being due to Ia antibodies. No change in expression of the private specificities was seen. On the other hand, during the later passages (239-258) a number of qualitative differences were seen, the most remarkbale being the loss of H-2.33 and gain of H-2.4, 31. The overall serological pattern of cells resembled that of H-2d rather than of H-2bp haplotyes and this was confirmed by absorption tests. The changes reported here may be due to alterations of repression and derepression pattern of the presumed multiple structural H-2 genes present in the genome.

Notizen: Cellular cytotoxicity experiments were done to test the potential of the extra H-2 antigenic specificities on the K36 spontaneous leukaemia as targets. In addition, experiments were performed to rule out the possibility that the target determinants could be normal cross-reacting alloantigens, e.g., T1, Qa, non-H-2 and previously undetected H-2 public specificities, which are more readily detectable on tumours than on normal cells.We used F1 hybrid mice, in which one parent was of the strain of origin of the tumour (K36) and the other parent of the B10 congenic series, i.e., (AKR x B10)F1. These cells were stimulated by lymphoid cells from other B10 congenic strains, B10.A and B10.D2, and tested against the test tumour K36 and several PHA blast controls. Several K36 sublines as well as a cloned line of K36 (K36.16) were used and significant cytotoxicity against. an H-2d-like target on these tumour cells was obtained. These data exclude the possibility of a corss-reactive alloantigen, e.g., undetected H-2 public specificity, or differentiation antigens. These results with the K36 tunour were consistent with our immunochemical studies (see Schmidt & Festenstein, 1980) and were confirmed and extended by cold target inhibition experiments. In these experiments, B10.BR cells were sensitized by B10.D2 lymphoid cells and tested against B10.D2 (51chromium-labelled PHA balsts). Two kinds of normal unlabelled lymph node suspensions as well as the K36.16 tumour cell suspension were used. Significant specific inhibition of between 19% and 40% was obtained using K36 and between 23% and 37% using B10.D2 (positive conrol). AKR cells (negative control) in contrast were unable to reduce the precentage specific cytotoxicity.Since it was already known that the H-2Kk gene products are missing from this tumour (Schmidt & Festenstein, 1980), it was of interest to test whether cytotoxic effectors directed against the H-2kk gene products were able to kill the K36 tumour. Accordingly, B10.D2 lymphoid cells were sensitized to B10.BR (C3H.0H) and B10.A targets, respectively, and tested against K36 and appropriate controls. Only weak killing was observed when sensitization was effected against the K end of the H-2k haplotype (i.e., using B10.A as the sensitizing cell) but strong and significant cytolysis was found when the sensitization was against the whole H-2k haplotype or against the H-2Dk gene product. These results were confirmed by cold target inhibition studies.These experiments provide further indications for the H-2d-like characteristics of these allodeterminants. We have already excluded some of the possible explanations for these findings (i.e., corss-reactions with H-2 and non-H-2 normal specificities). The cold target inhibition experiments rule out non-specific viral effects. Thus, we favour an alteration in regulatory genes leading to repression of the H-2Kk product and derepression of the H-2Dd product, but cannot formally rule out highly corss-reactive H-2-like virally encoded determinants.

Notizen: Alien H-2k-like antigens were found to be expressed by a methylcholanthrene induced tumour of BALB/c (H-2d) origin. H-2 specificities of the k haplotype were detected on this tumour by a variety of serological techiques, including 51Cr-release cytotoxicity, microradioassay and absorption. The antisera employed were conventional polyspecific alloantisera, typing sera with restricted specificty and monoclonal hybridoma-derived anti-H-2k antibodies.The tumour has a low expression of the private specificty 31, which characterizes Kd molecules, and does not seem to express the private specificities of Dd, Kk and Dk molecules. It appears to express predominantly alien H-2-like antigens which are very similar to but not identical with normal H-2k molecules.

Notizen: The immune response of SJL/J (H-25) mice to syngeneic reticulum cell sarcoma (RCS) tumour cells was investigated. The nature of tumour-associated antigens was examined by immunological and biochemical techniques. Reticulum cell sarcomas of different origins (spontaneous, transplantable and cultured cell lines) stimulate a strong syngeneic response measured by [3H]-thymidine incorporation. However, only the cell line were able to stimulate a syngeneic cell-mediated cytotoxic response. Further analysis of the syngeneic response revealed that RCS cells express inappropiate alloantigenic specificities on their surface. Thus, evidence is provided which demonstrates that RCS tumour cells carry antigens which cross-react with BALB/c (H-2d) and C57BL/6 (H-2b) alloantigens. The presence of inappropriate antigens in the in vitro lines was detected by cell-mediated cytotoxicity, cytotoxic antibody and immunofluorescence whereas antigens on in vivo lines and spontaneous RCS were detected primarily by immunofluorescence. Conversely, H-25 expression on the in vitro lines was detected only by immunofluorescence whereas the H-25 of the in vivo lines were detected by cytotoxic antibody. By these assays inappropriate antigens were not detected on normal SJL/J lymphocytes. Immunochemical analysis of the inappropriate antigens by two-dimensional gel electrophoresis was performed with SJL/J anti-BALB/c and monospecific anti-H-2Dd sera. These sera precipitated 45,000 MW molecules from BALB/c lymph node cells and from SJL/J in vivo and in vitro RCS tumour which appeared very similar. The sera also precipitated molecules from SJL/J lymph node cells which resemble the BALB/c and RCS molecules. Thus, RCS tumours express inappropriate alloantigens which may be cryptic on normal cells. The biological significance of these tumour-associated antigens in vivo is discussed briefly.

Notizen: Serological and structural changes of surface markers involved in immune reactions may occur in human and murine tumour systems. Thus nine out of twenty-one human tumour cell lines and SV40-transformed fibroblasts differed from autologous lymphoblastoid cells or fibroblasts in their reactivity with HLA allonatisera. H-2 antigens isolated from the murine tumour cells 6C3HED and TP 1422, undergo structural changes. An alien HLA-B7 was detected in sera from two melanoma patients. The serologic activity on H-2 antigens was significantly increased in the serum and ascites fluid of tumour bearing mice. Additionally, human SV40-transformed fibroblatsts acquire receptors for monkey red blood cells and the murine lymphosarcoma cells 6C3HED express receptors for sheep red blood cells.

Notizen: Morphological studies of metaphase chromosomes were done with rats derived from the BIL/1 strain, which has genes affecting growth and reproduction linked to the major histocompatibility complex by conventional Giemsa-trypsin staining and the results were compared to rat strains not carrying these defects. A subterminal-terminal centromeric polymorphism was detected in chromosome 3 [del 3 (pter → cent)] among the strains studied. Comparison of the G-banded karotypes of the rats carrying the defects with the karotypes of the otsher strains did not reveal any gross chromosomal abnormalities.

Notizen: C4-coated Ch(a+) red blood cells (RBC) were used as indicator cells in a serum inhibition reaction of anti-Cha, for the determination of the Ch group of serum. This serological study, combined with electrophoretic studies of C4 in a family material, showed that the C4M haplotype product was associated with partial inhibition of anti-Cha.

Notizen: Individuals with selective absence of IgG1 and IgG2 were discovered by testing for allotypes and isotypes of the respective sub-classes. These individuals were homozygous for sub-class deleted Gm-Am haplotypes, as shown by allotype studies in two families (Gm-;…;g;A2ml/Gm-;n;b;A2ml and Gm-;n;b;A2ml/Gm-;…;b;A2ml) and by a population study of New Guineans (Gm fa;-;b;A2m2). The individuals with IgGl sub-class deficiency showed elevation of IgG2, IgG4 and in particular of IgG3.Gene deletion can result from unequal crossing over which renders a complementary chromosome with a duplication of a sub-class gene. In one family, duplication of γ3 genes was observed to have happened in one of a twin pair. Quantitation of sub-classes in families with γ1- and with γ3-duplicated haplotypes did not show increased levels of the gene involved.

Notizen: A hybrid cell line R3/41-10 was produced by fusing a mouse myeloma line with spleen cells from CBA/H mice which had previously been immunized with human peripheral blood lymphocytes (PBL) obtained from a single donor. Hybrid secreting antibodies to lymphocyte antigens were detected by assaying the culture supernatant for antibody binding to the donor PBL. Those lymphocytes binding more than ten times the background were cloned in soft agar and transferred to micro-culture plates (Limbro). Antibodies from some wells lost their binding activity. Antibodies from other clones, although retaining their binding capacity, were multispecific in cytotoxicity experiments, killing all the lympoid cells from a panel of normal donors; yet a third kind gave specific limited reactions. Supernatants from clones R3/41-10 and R3/41-13 gave concordant cytotoxic reactions killing 20-40% of the PBL. There was no cytotoxicity of Ig+ cells (B cells). Lymphocytes from seven out of fourteen normal donors reacted with the antibody. The specificity of the antibody produced by clone R3/41-10 was confirmed by absorption studies.The monoclonal antibodies (Mc+b) described in this communication are shown to react with a subpopulation of T lymphocytes of some individuals and not others, suggesting that they are detecting a polymorphic system of alloantigens like the Ly system in the mouse, provisionally designated HT-Ly. l.

Notizen: Subcutaneous (s.c.) immunization of mice with viable allogeneic H-2 compatible spleen cells can induce a persistent state of delayed type hypersensitivity (DTH) to minor histocompatibility (H) antigens which can be evaluated with a footpad swelling assay. The importance of H-2 compatibility of the injected spleen cells with the recipient for (1) the elicitation of the DTH-reaction, (2) the induction of DTH-related effector and memory T cells and (3) the activation of T memory cells was examined with congenic mouse strains. Spleen cells sharing either the K or D region of the H-2 complex with the recipient could elicit strong DTH-reactions to minor H antigens, though somewhat less than did fully H-2 compatible allogeneic spleen cells. H-2 incompatible cells or cells only sharing I-region coded antigens elicited relatively weak, though significant, DTH-reactivity to the minor H antigens. Similar H-2 requirements for recipient mice were demonstrated in the immune lymphocyte transfer assay. Optimal induction of primary DTH and DTH-related T memory cells for minor H antigens also required H-2 identity of the immunizing cells and the recipient. To some extent, H-2 incompatible cells were able to induce primary DTH-reactivity and memory for minor H-antigens. The secondary DTH-reactivity was not or slightly dependent on the H-2 haplotype of the cells used for booster injection.It is concluded that the DTH-related effector cells are restricted in their recongnition of minor H antigens by K- or D-region-coded antigens. Presumably, macrophage processing of the allogeneic spleen cells after primary and secondary immunization accounts for the capacity of the minor H antigens to activate unprimed T cells and memory T cells when these antigens are presented on H-2 incompatible cells.

Notizen: Difference in the amounts of H-2K.5 antigens present on erythrocytes was observed, using quantitative absorption method, among several B10 congenic strains. However, no such variation was seen on the lymphocytes and lung cells. The variability in the amount of this antigen on the erythrocyte surface was primarily dependent on the haplotype of the H-2K end in the B10 recombinant strains examined. This suggests that the regulator of H-2 expression on erythrocytes is in this region. Further genetic analysis confirmed that this regulation functions in the cis-position. Finally, the H-2K.5 antigenic activity was expressed on the reticulocytes of all strains tested, but appeared lower in the type 2 group indicating that the regulation begins early in erythropoietic differentiation and eventually results in a complete loss of detectable activity.

Notizen: A two-step procedure employing gel filtration and anion exchange chromatography has been utilized to isolate LMW immunoglobulin from the horned shark, Heterodontus francisci. Light chains obtained by complete reduction and alkylation of the parent protein have been compared by several analytical techniques. Amino acid composition data implies a limited degree of variation in the light chains isolated from individual animals. Polyacrylamide gel electrophoresis of the CNBr digests of the light chains reveal indistinguishable banding profiles of the major peptides. Isoelectric focusing indicates limited heterogeneity in the light chain spectrotype and identity in the pI of the majority of bands detectable by staining. The suggested degree of structural similarity in the light chains of this phylogenetically primitive shark is discussed in terms of the evolutionary position of the species and current theories concerning the origins of structural diversity in immunoglobulins.

Notizen: The mechanism of neonatally induced transplantation tolerance was studied in two mouse strain combinations involving differences at the D region of the H-2 complex only or at the same D region plus I-J subregion (including I-E, I-C, S and G regions). In the strain combination with the H-2D difference only, cells from tolerant mice proliferated markedly in the MLR assay when incubated with antigens tolerated in vivo, whereas the MLR reactions were negative in the combination with D plus I-J region disparities. In the latter combination cells from tolerant mice also did not respond to third-party antigens and their incubation with the tolerated antigens led to the suppression of cell proliferation. This non-specific suppression was absent in cells from tolerant mice in the strain combination, which differed in I-C, S, G and D alloantigens. Specific suppressor cells, which inhibited the development of cytotoxic cells, were demonstrated in tolerant mice of both strain combinations. The results show that, in addition to the specific suppressor cells induced by H-2K or H-2D alloantigens, non-specific suppressor cells induced by the I-J region disparity that may regulate the resultant activity against H-2D (and probably also H-2K) alloantigens are involved in transplantation tolerance.

Notizen: We have used a combined serologic and structural approach to study the distribution of I-region associated (Ia) antigens in nine strains of inbred and partially inbred guinea-pigs. All of the inbred strains studied with the exception of strain 2 animals were found to share one or more I-subregions with inbred strain 13 animals. The BIOAD, R9, OM3, and BIOAC strains have the same I-region as strain 13 animals; the B/Lac strain has two subregions in common with strain 13, while the BIOB strain has a single subregion in common with strain 13. The availability of a number of different guinea-pig strains with well characterized major histocompatibility complexes should facilitate the continuing use of this species in studies of immunogenetics, transplantation, and tumour immunology.

Notizen: Two-way and one-way mixed lymphocyte cultures (MLC) with Chinese Hamster peripheral blood lymphocytes were performed. Reduction of the culture surface and addition of 2-mercaptoethanol to the culture medium gives a marked enhancement of the reactivity of low numbers of lymphocytes in the MLC. Genetic analysis of the mixed lymphocyte reactivity (MLR) in the animals studied so far indicates that three or four different alleles are involved. Howerver, not all phenotypes could be traced. The samples of different stocks of Chinese Hamsters in Europe and the United States did not all share the same alleles and the samples were also different as regards the relative frequency of phenotypes. For DNA inhibition in stimulating cells mitomycin C treatment is less suitable than irradiation. Skin graft survival time was longer in animals which showed no MLR than in animals with a positive MLR.

Notizen: The genetic basis of the restriction imposed on T cell mediating acquired antimicrobial resistance and delayed-type hypersensitivity (DTH) to Listeria in the rat was investigated. Sharing of MHC-coded genes between donors of sensitized T cells and antigen-stimulated recipients was both necessary and sufficient for efficient transfer of both resistance and DTH. Evidence to support this assumption was derived from experiments involving allogeneic transfers within major histocompatibility complex (MHC)-compatible strains and across MHC-barriers. Further support came from linkage studies with backcrossed rats and with the progeny of F1 rats mated with an unrelated strain. An unexpected difference in the compatibility requirements for effective transfer of DTH and resistance was noted in experiments involving the BI strain (formerly called B3). Thus, while B-region compatibility was obligatory for expression of DTH in recipients of sensitized T cells, considerable levels of protection could be transferred to either A-region or B-region compatible hosts.

Notizen: The antibody response in mice to type I calf skin collagen is quantitatively determined by genes which map to the I region of the H-2 histocompatibility complex. The use of H-2 recombinant B10 congenic strains of mice reveals that a gene in the IA subregion and a gene to the right of the IA subregion affect responsivenss. To examine complementation patterns in the antibody response to collagen, five B10 congenic strains, each bearing an independent H-2 haplotype, were intercrossed to obtain nine hybrid strains heterozygous at the H-2 locus. In five combinations heterozygous progeny produced significantly greater antibody responses than those observed for the homozygous parental strains. Two low responder haplotypes, H-2k and H-2d, were shown to be qualitatively different. Mice of these haplotypes show a different dose-response pattern and a different phenotypic pattern of inheritance with respect to the high responder H-2b haplotype. Complementation effects found in F1 hybrid mice derived from H-2 recombinant parental strains indicate that high responsiveness, controlled by an IAb subregion gene, can be influenced by an interaction between an IAk subregion gene and an ICd subregion gene on different chromosomes. These data are consistent with the possibility that there exist two or more I region genes that have distinct functions and can interact to affect the levels of immune responsiveness.

Notizen: A study of Gm allotypes in a Caucasoid family with hypogammaglobulinaemic probands, showed qualitative (unexpected or lacking Gm allotypes) and quantitative (increased or decreased Gm contents) abnormalities in many relatives. Part of these observations can be most probably accounted for by inheritance of a Gm1,17;5,28 haplotype, not described in Caucasians yet, and by an in vivo expression of latent Gm genes.

Notizen: α-L-Fucosyltransferases were demonstrated in human saliva which catalyze the transfer of L-fucose from GDP-L-[14C]-fucose to oligosaccharides from human milk. An α-(1→4)-L-fucosyltransferase that synthesizes lacto-N-fucopentaose II and lacto-N-difucohexaose I from lacto-N-tetraose and lacto-N-fucopentaose I, respectively, was detected in saliva samples of Le(a-b+) secretors and Le(a + b-) non-secretors in which Lea substance was secreted. This enzyme activity was demonstrable neither in saliva samples of Le(a-b-) secretors nor non-secretors. An α-(1→2)-L-fucosyltransferase, that synthesizes lacto-N-fucopentaose I from lacto-N-tetraose, was detected in saliva samples from Le(a-b+) secretors which secreted H and Leb substances and from Le(a-b-) secretors which secreted only H substance. An α-(1→3)-L-fucosyltransferase was present in all saliva samples of different ABO and Lewis blood groups, irrespective of their ABH secretor status of the donors. The fucosyltransferases in saliva were activated by Mn++ or Mg++ ions, and were inhibited by ATP, GTP and EDTA. They had a broad pH optimun between pH 5.0 and 6.5.

Notizen: Genes in the D region of the murine major histocompatibility complex, H-2, confer resistance to radiation-induced leukaemia virus. H-2D gene control appears to ensue at a step subsequent to virus infection, since elimination of virus infected cells does not become apparent until 3-5 weeks after virus infection. Nonetheless, almost immediately after virus infection, expression of H-2D-coded antigens is markedly elevated on the surface of thymocytes from resistant (H-2Dd) but not susceptible mice (H-2Ds or H-2Dq). This increased H-2D antigen expression triggers a vigorous cell-mediated immune response which probably plays a key role in resistance to leukaemia via elimination of virus-infected cells. A hypothesis is put forth to explain the induction of increased sythesis and expression of H-2D antigens. This hypothesis postulates that the oncogenic segment of RadLV bears a close resemblance to H-2.4, the private specificity for H-2Dd, allowing it to integrate at or near the H-2Dd murine gene. Subsequent to integration, the rates of transcription and translation are altered with a resulting increase in cell surface antigen expression. Other possibilities are also discussed.

Notizen: When testing the serum of an individual anti-H-2 immunized mouse (B10 x A.SW)F1 anti-B10.M by the routine micro-lymphocytotoxicity test on lymph-node cells, unexpected antibodies were found. The most striking finding was that after absorption of anti-H-2.8 antibodies with B10.A(2R) (Kk) cells, antibodies remained which reacted with AKR, B10.AKM and B10.A V + mice while B10.A V-, B10.BR and C3H mice were negative. While all these strains share the Kk allele, only the positively reacting strains express high titres of infectious RNA turnover viruses. Unexpected reactions were observed also with H-2d, H-2j and H-2r cells and absorption experiments indicated two or three antibody populations.These reactions could be interpreted by two different possibilities: (1) anti-H-2 antibodies react with virus-altered H-2 structures; and (2) antiviral antibodies react with H-2 structures complexed with viruses. These possibilities should be taken into account when H-2 sera are tested on tumour or virus-infected cells.

Notizen: Cytostatis of the H-2d tumour LSTRA by H-2-restricted effector lymphocytes was inhibited by antisera against H-2.4 and H-2.31 but not by antisera against public specificities or non-H-2 antigens. The unexpected reaction of the same effector cells against Gardner tumour (H-2k) was also shown to be inhibited by a combination of antisera against H-2.4 and H-2.31 but not by each antiserum used separately. The inhibitory capacity of these antisera was removed by absorption with B10.D2 but not with B10 lymphocytes. This indicated the presence of H-2d-like specificities on gardner tumour which could function as self-recognition structures in an H-2Kd and H-2Dd restricted system.

Notizen: The specificity of he corss-killing exerted by cytotoxic T lymphocytes (CTL's) generated against H-2 region products was investigated in a 51Cr release assay on a panel of target cells from a number of different H-2 haplotypes.Their pattern of reaction shows that: (1) The target cells, expressing public specificities (H-2.28, H-2.1, H-2.5, H-2.3 or H-2.8) which should, theoretically, be recognized by the CTL's were killed, while those expressing no public specificities, recognized according to the H-2 chart by the CTL's wer not killed. (2) The CTL's generated against the H-2.28 specificity expressed on the D region products cross react with target cells expressing this specificity in their K region products and vice versa. The same phenomenon was observed with the H-2.1 specificity. These results provide evidence that public specificities are targets for CTL's. Antiserum reacting against the public specificity recognized by the CTL's was found to block the cross-killing, however, to the same extent as antisera directed against any specificity recognized by the CTL's was found to block the cross-killing, however, to the same extent as antisera directed against any specificity (private or public) expressed on the same molecule as the target determinant. Finally both the inhibition studies using anti-H-2 antisera and the direct cytotoxic assays showed that the public specificities of the H-2.28 family carried by the H-2.D and H-2.L molecules were recognized by different subpopulations of CTL's.

Notizen: Abstract. The transmission of Yersinia ruckeri has been investigated in steelhead trout using asymptomatic carriers of the causative bacterium of enteric redmouth disease. It was found that unstressed carrier fish did not transmit the bacterium to recipient fish to cause either an epizootic or produce new carrier fish. However, when the carriers were stressed with heat, the bacterium was transmitted from the carrier to recipient fish producing a lower intestinal carrier state but no deaths. Examination of experimentally infected fish to determine the number of carriers among the survivors indicated that the frequency varied as a function of time following infection. When immunized fish, were challenged with Y. ruckeri they became temporary carriers of the bacterium for up to 3 days; but were not able to transmit the infection to healthy recipient fish.

Notizen: Book reviewed in this article: Diversity and Adaptation in Fish Behaviour. By M. H. A. Keenleyside. Exotic Species in Mariculture. By R. Mann. Developments in Marine Biology. Vol. 1. Toxic Dinoflagellate Blooms. Ed. by D. L. Taylor and H. H. Seliger Phytoplankton and the Fish Kills in Loch Striven. Ed by P. Tett.

Notizen: Abstract. Atkinsiella hamanaensis sp. nov. a marine mastigomycete isolated from ova of the mangrove crab, Scylla serrata (Forsskål), is described and illustrated. The fungus grew over a temperature range of 15–32°C, with an optimum of 29–32°C. Its growth was observed in peptone-yeast extract glucose broth containing 1–5% NaCl, with optimum growth at 2–3% NaCl concentration. At 6% or more NaCl concentration, growth was inhibited. Its pH tolerance ranged from 4 to 9.

Notizen: Abstract. Flagellates infesting the gills of freshwater fish are reviewed and Cryptobia branchialis Nie is differentiated from them. Gill filaments of two species of fish heavily infested with C. branchialis were examined with the transmission electron microscope. The flagellates are attached to the surface of the gill epithelium by their recurrent flagella. A prominent ridge on the flagellar membrane contacts the host cell membrane, forming a zone of gap junction and leaving a space of about 10 nm between the two membranes. The flagella do not penetrate into the epithelial cells. No other modifications of the cell membrane could be detected and the host cell reveals no signs of damage. Assumptions on the pathogenicity of C. branchialis are discussed in view of ultrastructural findings, which suggest that this flagellate is non-pathogenic and ectocommensal.

Notizen: Abstract. This paper reports for the first time the application of the direct negative staining technique for the rapid confirmation of the viral nature of blenny‘piscine erythrocytic necrosis (PEN). The virus in negatively stained preparations has an internal core structure which appears in exact opposite contrast to that seen in thin sectioned virus. The resultant images from both negative staining and thin sectioning suggest that the organization of the core is unique amongst intracytoplasmic deoxyriboviruses (ICDVs).

Notizen: Abstract. Electron microscopical and histopathological investigations were carried out on the anterior kidney often 1-year-old farmed rainbow trout naturally infected with the external signs of viral haemorrhagic septicaemia (VHS). The histopathology of the liver and the haematology of peripheral blood were also examined. The presumptive virus of VHS was directly demonstrated for the first time in the anterior kidney.All diseased fishes showed necrotic changes in the kidney and the liver. The melano-macrophage centres of the anterior kidney showed a partial destruction accompanied with a loss of melanin. A progressive destruction of the erythrocytes was observed. The liver exhibited moderate to extensive necrosis (karyorrhexis and karyolysis with lymphocytic invasion in the necrotic areas). Neither intranuclear nor intracytoplasmatic inclusions were observed. In the peripheral blood diverse stages of erythrocytic degeneration, amitoses of the erythrocytes and a five-fold increase in lymphocyte numbers were observed.

Notizen: Abstract. Perfusion fixation of trout kidney with picric-acid: formaldehyde: acetic acid: ethanol mixture was compared with other methods as a fixative for staining with the alcian-blue PAS technique. Perfusion fixation gave conspicuously better results for light microscopical purposes.

Notizen: Abstract. Experimental infection of artificially-induced skin ulcers in rainbow trout Salmo gairdneri Richardson with an achromogenic strain of Aeromonas salmonicida induced a syndrome similar to that observed in cyprinid fishes. Sequential histological observations showed that the presence of the bacteria completely inhibited any migration of peripheral epidermal cells and subsequent under-running of the ulcer resulted in its expansion, as peripheral epidermis sloughed.

Notizen: Abstract. New methods were studied for the control of Ichthyophthirius muttifiliis Fouquet, one of the most serious parasites of fish. Totals of 272 fish, 354 I. Multifiliis trophozoites and 562,500 tomites were used in the screening of amprolium, ronidizole, and silver nitrate for parasite control. In addition, 1,465 trophozoites and 600 tomites were subjected to pulsed DC electrotherapy trials. Silver nitrate at 0·67 ppm was 100% effective in eradicating tomites, while ronidizole at 750 ppm gave 100% and 97% control of trophozoites and tomites, respectively. Amprolium and electrotherapy were both ineffective against trophozoites and tomites. Silver nitrate has promise as a safe and effective alternative to chemicals currently used to control I. multifiliis.

Notizen: Abstract. The prevalence and pathology of the pseudophyllidean cestode, Polyonchobothrium clarias in the gastrointestinal tract of the teleost, Clarias mossambi-cus have been studied. The cestodes were most abundant in the gall bladder where they caused pronounced nodular outgrowths in the mucosa and exhibited a gradual decrease in number distally towards the rectum. Generally the intensity of the infection due to P. clarias was higher in young fish than in old fish.

Notizen: Abstract. Experiments with a purified vitamin test diet on newly hatched Cirrhina mrigala Hamilton, a major carp of great piscicultural importance, are reported. Experimentally induced vitamin C deficiency for 330 days was followed by recovery treatment for 30 days. Retarded growth and greater mortality accompanied by structural deformities such as lordosis and scoliosis, increasing with prolonged deficiency, were recorded. Haematological and haematopoietic studies revealed hypochromic macrocytic anaemia accompanied by anisocytosis. Correlated studies on erythropoiesis revealed increased reticulocytes and an overall decrease in small lymphoid haemoblasts. Total and differential leucocyte counts revealed leucopenia and thrombopenia accompanied by an increase in neutrophils and eosinophils. Recovery treatment halted and in some cases even compensated these pathophysio-logical changes to some extent.

Notizen: Book reviewed in this article: Aquaculture-Institute of Biology Studies in Biology No. 106. By P. J. Reay. Immunity in Evolution. By J. J. Marchalonis

Notizen: Abstract. The 96-h medium tolerance limit of Furanace (nifurpirinol: P-7138) for channel catfish Ictalurus punctatus at 21°C was 0-94 mg/1. The most effective treatment level for channel catfish, experimentally infected with Aeromonas hydrophila, was 2mg/1 for 6.5 h. Furanace at 0.5 and 1.0 mg/1 in brain heart infusion agar reduced in vitro growth of A. hydrophila and at 2.0 mg/1 growth was completely

Notizen: Abstract. Starry flounder Platichthys stellatus (Pallas) from the Duwamish River estuary, which has received large volumes of pollutants in the past, and starry flounder from two relatively unpolluted reference areas were examined for pathological conditions. Gross and microscopic examination of starry flounders from the Duwamish River estuary revealed one or more of the following unusual liver characteristics in 92% (monthly range 85–100%) of the fish: liver discolouration, liver enlargement, severe fatty vacuolation of the hepatocytes, centrolobular cellular degeneration and necrosis of hepatocytes, megalocytic hepatocytes, proliferating perivascular fibroblasts displacing adjacent liver parenchyma, hepatic structural disarray and increased basophilia of hepatocytes in localized areas. Livers of starry flounders, usually spent spawners, from one of the reference areas also had one or more of these characteristics, although at a much lower frequency (〈13%), and these included liver discolouration, increased fatty vacuolation of the hepatocytes and hepatic structural disarray.

Notizen: Abstract. Cultured turbot Scophthalmus maximus (L.) from three farms at Hunterston, Scotland were found to be infected with Haemogregarina sachai. Up to 6% of some populations were affected with gross tumorous lesions of the musculature and viscera. The lesions consisted of necrotic tissue with a caseous centre. Histologically, there was an accumulation of parasitized reticulo-endothelial cells, cell debris and pyknotic nuclei. Parasitaemias of up to 36% of all blood cell types were observed. The majority of infected cells were neutrophils and monocytes. Infected leucocytes were hypertrophied. There was an erythrocytopenia of infected fish with increases in erythroblasts, monocytes and neutrophils but a decrease in thrombocytes and lymphocytes. Transmission experiments between fish were unsuccessful.

Notizen: Abstract. Serum thyroxine (T4) and triiodothyroninc (T3) levels were significantly lower in fish fed a high PCB diet (500 mg/kg) in comparison to those fed the control diet. The differences in serum thyroid hormone levels between the PCB-fed group and those fed the control diet were not evident after 14 days food deprivation nor after testosterone-injection. The low PCB diet (50 mg/kg) was without effect on serum T4 and T3 levels regardless of the treatment. Trout fed the highest Mirex diet (50 mg/kg) had T3 levels which were significantly lower and a T4 T3 ratio which T3was still evident after 14 days food deprivation but after testosterone administration the serum T3 levels in the control fish had fallen to levels similar to those in the Mirex-fed group. The low Mires diet (5 mg/kg) was without effect on serum T4 and T3 levels regardless of treatment. Serum T4 and T3 levels in fish fed a mixed PCB (50 mg/kg) and Mirex (5 mg/kg) diet were not significantly different from controls but T3 levels were significantly higher than in control fish after 14 days food deprivation and subsequent testosterone administration. There were no appearence difference in anterior pituitary or thyroid histology between the different groups. The hepatosomatic index (HSI) in PCB-fed trout was significantly larger than in controls in the fed groups, but not after 14 days food deprivation and subsequent testosterone administration. There was a five-fold difference in carcass PCB hioaccumulation between fish fed the two PCB-conlaminated diets, despite a 10-fold difference in dietary levels of the organochlorine. Similarly despite 10-fold differences in dietary Mirex levels in fish fed the 5 and 50 mg/kg and the 50 and 500 mg/kg diets, there were only 4–56–and 1–35-fold increases in carcass Mirex content, respectively. Trout fed the mixed Mirex-PCB diet had PCB levels of only 60.2% of those fed comparable levels of PCB alone (50 mg/kg) but contained similar levels of Mirex to those fed Mirex alone (a mg/kg).

Notizen: Abstract. An autoradiographic study of the various types of neoplastic lesions in the skin of Pacific English sole Parophrys vetulus Girard was performed using peritoneally injected tritiated thymidine. The epidermis offish is mitotically active at all cell levels, the most important being the supra-basal layer. In the hyperplastically growing epidermis of the English sole covering connective tissue nodules and in the papillating epidermis prior to formation of enlarged ovoid cells (X-cells), the supra-basal layer also shows the greatest degree of labelling. Parallel to the appearance of these X-cells, which are characteristic of the skin tumours in Pacific flatfish, the distribution of DNA-synthesizing cells is altered. In the tumour areas which are more or less filled with X-cells the ‘envelope cells’ are labelled throughout with a uniform frequency. On the other hand, the directly adjacent X-cell free areas are labelled with a greater than average frequency.

Notizen: Abstract. A comparative diagnostic study was conducted on goldfish, Carassius auratus (L.), with a cutaneous ulcerative disease from five locations in the United States and one each in England and Japan. Fish were examined for parasites, viruses and bacteria. Fish from all locations examine d were infested by ectoparasites; no single parasite species was common to all locations. No virus-associated cytopathology was observed in fathead minnow (FHM) or adult goldfish (CAR) monolayer cell cultures inoculated with homogenates of cutaneous lesions, kidneys and livers from diseased fish. The only bacterium cultured from fish from all locations was an atypical, often late-pigmenting strain of Aeromonas salmonicida. This organism was isolated from 64 of 83 (77%) of the total lesions cultured and was most prevalent in early lesions. A second commonly isolated organism was Aeromonas hydrophila, which was cultured from fish at four of the seven locations and from 28 (34%) of the total lesions cultured. A. hydrophila was most prevalent in terminal lesions. From these studies it was concluded that A. salmonicida was the probable cause of ulcers noted In the cases examined an d that A. hydrophila was a secondary invader.

Notizen: Abstract. Infection experiments were conducted to determine the primary aetiology of an ulcerative disease in goldish, Carassius auratus (L.). Goldfish were exposed to atypical Aeromonas salmonicida and A. hydrophila, previousl y isolated from cutaneous ulcers, and to 04 5 μm filtrates of cutaneous ulcers and kidneys from diseased fish. Fish were exposed to each preparation by intraperitoneal, intramuscular or subcutaneous injection and by a method in which a small patch of scales was removed from each side of the fish and the inoculums applied. Most of the fish injected with A. salmonicida died without developing coetaneous ulcers; however, ulcers were induced in five of the ten fish exposed by the scale removal technique. Exposure to ultra filtrates or A. hydrophila did not result in consistent ulcer formation o r death. Additional experiments showed that a 30-min exposure of goldfish, without prior treatment, to water containing 3 × 105 colony forming units (cfu/ml) of A. salnumicida was sufficient to produce cutaneous lesions in nine often fish exposed. Multiple lesions were produced in most fish and A. salmonicida was consistently recovered. Fish exposed by similar waterborne challenge s to 6·2 × 106 cfu/ml of A. hydrophila or to 7·2 × 106 cfu/ml of another lesion isolate identified as a member of the A. hydrophila complex produced no lesions, eve n when scales were removed. The studies demonstrate that atypical A. salmonicida can initiate cutaneous lesion s characteristic of ulcerative disease, while A. hydrophila and an A. hydrophila complex organism cannot.

Notizen: Abstract. An ‘oedema syndrome’ caused mortality of 55·5% in brood brown trout approximately 2 months after spawning. The pathology and clinical biochemistry of the condition are described. It is suggested that the severe hypoproteinaemia and anaemia were due mainly to the traditional reduction in feed levels prior to spawning at a time when the water temperatures were abnormally high. It is also suggested, however, that the syndrome would not have occurred if the fish had not developed an aeromonad infection prior to stripping. Concurrent fungal infection and subsequent extremely low water temperatures are also thought to have played a role.

Notizen: Book reviewed in this article: Seawater Aquariums: The Captive Environment. By S. Spotte. Fish Cultivation in Sewage Stabilization Pond Effluent. By P. Edwards The Use and Potential of Aquatic Species for Wastewater Treatment. Publication No. 65. Appendix B. The Environmental Requirements of Fish. By J. Colt, S. Mitchell, G. Tchobanoglais & A. Knight. Marine Maladies? Worms, Germs and Other Symbionts from the Northern Gulf of Mexico. By R. M. Overstreet

Notizen: Abstract. Antigen localization studies in the mirror carp Cyprinus carpio were carried out using a soluble antigen, human gamma globulin and a cellular bacterial antigen, Aeromonas salmonicida. Antigen was detected using direct single layer and indirect double layer immunofluorescent techniques, respectively. A study of the spleen, pronephros, mesonephros, liver and thymus at various times after administration of the antigen showed that fluorescence occurs mainly in the ellipsoids of the spleen and in the haemopoietic parenchyma of the pronephros and mesonephros and reaches its greatest intensity when antibody titres are relatively high. A direct comparison with antigen localization in the clawed toad Xenopus laevis, has enabled us to analyse possible evolutionary trends towards the complex dendritic retention of antigen seen in the higher vertebrates.

Notizen: Abstract. The circulating haemocytes of the tanner crab, Chionocetes bairdi, were studied and classified as hyalinocytes, intermediate granulocytes and eosinophilic granulocytes. Differential haemocyte counts of normal crabs and crabs infected with an apparently pathogenic unnamed ascomycete fungus revealed a shift in the hyalinocyte-granulocyte ratio in infected crabs. There was a highly significant increase in the percentage of granulocytes and a corresponding decrease in hyalinocytes in infected crabs. This shift was thought to be caused by a marked increase in the relative number of eosinophilic gianulocytes. There was also a highly significant statistical relationship between the severity of the internal fungus infection, established by histopathological analysis of the major organ systems, and the increased number of circulating eosinophilic granulocytes. The results of the study indicate that differential haemocyte counts may be a valuable tool in monitoring the health of C. bairdi populations.

Notizen: Abstract. Histopathological studies on natural and experimental infections of nineteen microsporidian species from fishes distinguished two types of tissue reactions.The first type is characteristic of infections with xenoma-inducing microsporidian species and comprises three successive stages: a weakly reactive stage, a productive stage with the formation of granulomas and a stage of granuloma involution. Following the first stage, tissue reactions are directed towards the isolation of the parasite and result in its complete elimination and host tissue repair. The extent of pathological changes probably depends on the number of parasite cells which initiated the infection.The second type is represented by Pleistophora species infecting muscles or oocytes. Host tissue reaction is surprisingly slight during the schizogony and sporogony and does not tend to isolate the invaded muscle fibres. A slight lymphocytic infiltration of myosepta indicates the first stage of tissue reaction. The tissue reaction only reaches the productive stage when mature spores completely fill the contents of the infected muscle fibre. A thick wall of fibroblasts may be formed to demarcate the parasite mass as soon as it undergoes necrotic changes. The extent of pathological changes probably depends on the ability of early developmental stages of the parasite to spread the infection within the host.In both types of tissue reaction, the spores are destroyed by complete digestion within host phagocytic cells.

Notizen: Abstract. A vacuum degassing apparatus was used to subject channel catfish Ictalurus punctatus (Rafinesque) to sublethal hypoxia for 24, 48 or 72 h. Spleen, liver, gills, anterior kidney and posterior kidney from the catfish exhibited histopathological changes that included necrosis, hyperaemia, oedema, haemorrhage, hyperplasia and hypertrophy. Of the organs examined, gills were most affected by the hypoxic conditions. After 5 days of reacclimatization to normoxic conditions, channel catfish continued to exhibit histopathological effects of prolonged hypoxia.

Notizen: Abstract. A normally free-living hymenostomatid ciliate was found to be the cause of heavy infections of gills, viscera and body muscle in Atlantic and Pacific marine fishes kept in the New York Aquarium. Of special interest is the fact that the parasites, usually with food vacuoles filled with blood cells and cellular debris, caused extensive damage to the body muscle; the other infected organs showed no characteristic tissue reactions. The immediate cause of death is attributed to the exceptional load of the parasite on the gills, thus interfering with normal respiratory function.The ciliate, isolated from a recent outbreak, is described in detail, and shows many characteristics of Uronema marinum Dujardin, a species that has been reported as free-living in salt water; the form reported here differs from the type and other members included in this species, by the presence of less than three rows of bipolar meridians. This facultative parasite is described in detail and compared with other genera.

Notizen: Abstract. Bacteriologically sterile filtrates of ‘tumorous’ fish elicited formation of ‘tumours’in vivo which were morphologically indistinguishable from those described by Wessing & Bargen (1959). Attempts to isolate the putative aetiologic ‘Virus’ in primary and established cell cultures failed. When the lesions were examined for other compatible aetiologies, acid-fast stains revealed acid-fast bacilli and cultures grew Mycobacterium fortuitum. It is suggested that the ‘kidney tumour virus’ is actually soluble mycobacterial cell-fractions capable of eliciting tubercle formation in vivo.

Notizen: Book reviewed in this article: Sea microbes. By J. M. Siebueth. Farming the flat oysters of the genus Ostrea. By P. Korringa. Animal Tissue Techniques. By G. L. Humason. 4th Edn. Reef Fishes of the Sea of Cortez: The Rocky-Shore Fishes of the Gulf of California. By D. A. Thomson, L. T. Findley and A. N. Kerstitch. Tissues and Organs Text-Atlas of Scanning Electron Microscopy. By R. G. Kessel and R. H. Kardon.

Notizen: Abstract. Pleistophora typicalis Gurley, 1893, a parasite of the striated muscle of Myoxocephalus scorpius has been re-examined at light- and electron-microscope levels. Foci of infection were almost exclusively found in the ventral body wall where a mixture of schizonts, sporonts and masses of pansporoblasts lay in direct contact with unaltered host myofibrils and mitochondria.All stages were surrounded by a thick (0-5 μm) amorphous coat, external to the plasmalemma. In schizonts this was traversed by channels passing from the plasma-lemma to a layer of vesicles in contact with the muscle. This coat became modified as a pansporoblast envelope surrounding the mature spores: at first a layer of fine granules was laid down about mid-way across the previously amorphous coat, while the channels disappeared. At the time when the sporogonial plasmodium retracted away from the coat to produce the pansporoblast cavity, the layer of the coat within the granules disintegrated into spherical vesicles. The pansporoblast envelope around the mature spores was composed of two or three layers of different electron density, including one which was strongly electron dense. The pansporoblast envelope as defined here corresponds in function, but not necessarily in origin, to the sporophorous vesicle that encloses the spores in other pansporoblastic microsporidia.Schizonts had an extensive system of smooth endoplasmic reticulum, composed of expanded vesicles and divided by plasmotomy into smaller multinucleate segments. The endoplasmic reticulum of sporogonial stages was comprised of a network of fine channels, parallel arrays of flat cysternae and close-packed stacks of membranes. The sporogonial plasmodium divided stepwise through smaller multinucleate segments into uninucleate sporoblasts. Nuclei were isolated throughout development.Two types of sporonts were recognized: large sporonts gave rise to pansporoblasts containing up to 200 microspores. Macrospore pansporoblasts always contained eight macrospores. Microspores measured 4–4 × 2·3 μm (fresh) and were probably uninucleate. Macrospores measured 7·5 × 3·0 μm (fresh) and were probably binucleate.Another microsporidium, found in the muscles of Blennius pholis, which had been attributed to the same species by Thélohan (1895), was distinguished on the basis of spore size (microspores 3·9 × 2·3 μm and macrospores 7·7 × 3·8 μm fresh) and by the fact that electron dense components were laid down in the pansporoblast wall early in sporogony. The species was named Pleistophora littoralis n.sp.

Notizen: Abstract. A bacterium resembling Aeromonas salmonicida and determined to be the aetiologic agent of a cutaneous ulcerative disease in goldfish Carassius auratus (L.) was further characterized in this study.Forty-five isolates of the bacterium (43 from the United States and one each position (moles % guanine plus cytosine) and DNA homology. The bacteriological atypical A. salmonicida previously described. Several important biochemical characteristics distinguished the goldfish isolates from typical A. salmanicida, but the DNA binding experiments indicated a high degree of relatedness between the goldfish isolates and typical A. salmonicida strains.

Notizen: Abstract. A viral agent (13p2), isolated from clinically normal juvenile American oysters (Crassostrea virginica) and characterized as a new serotype of reovirus, was tested to determine if it could replicate and produce disease in experimental juvenile oysters. Because the virus replicated well in the bluegill fry (BF-2) fish cell line, fingerling bluegills Lepomis macrochirus were included in the pathogenicity experiments. Exposure of oysters to the 13p2 virus in ambient seawater resulted in no significant mortality and no increased virus titres or histological lesions. Virus particles were not observed in tissues of exposed oysters when examined with transmission electron microscopy (TEM). Intraperitoneal inoculation of the 13p2 virus into bluegills resulted in 44% mortality associated with a focal necrotic hepatitis. Evidence of virus replication, when evaluated histological or by virus titration, was observed in 94% of 32 inoculated fish. Samples of infected livers examined with TEM revealed typical cytoplasmic arrays of 13p2 virus particles in affected hepatocytes. Rising virus titres and hepatic lesions also occurred in bluegills exposed to water containing the 13p2 virus. These results indicated the natural host of the 13p2 virus was not the American oyster, but that it was a significant pathogen for at least one fish species.

Notizen: Books reviewed in this article:Advances in Aquaculture. Papers presented at the FAO Technical Conference on Aquaculture, Kyoto, Japan, 26th May-2nd June 1976. Ed. by T. V. R. Pillav & W. A. Dill.Methods for Assessment of Fish Production in Fresh Waters. Ed. by T. B. Bagenal.

Notizen: Abstract. Ultrastructural observations on a possible aetiological agent and tissue changes in a previously described disease of hatchery reared larval oysters Crassostrea gigas Thünberg are reported. Spherical bodies, implicated in gastro-intestinal impaction, show a strong resemblance to the marine fungus, Hyalochlorella marina. Similarities include a thick outer wall structure, plate-like mitochondria and dense cytoplasm with small vacuoles.The dense fascicular masses, previously observed histologically at epithelial surfaces, consist of accumulations of irregular electron opaque bodies. Large accumulations of these bodies appear to exacerbate tissue damage and prevent normal velar function. In comparison with the reported virus-like particles in this disease, these dense inclusions and associated lesions are more frequently observed and thus seem to be more important in the disease process.Notes on normal epithelial surfaces of larval Crassostrea spp. are included.

Notizen: Abstract. Sphaerospora angulata Fujita, 1912, has been identified as a frequent renal parasite of cultured carp populations in Hungary. Its prevalence ranged from 10 to 50% in the pond farms surveyed. The youngest carp infected were 21/2–3 months old. Infections were not common in one-summer fish and tended to become less frequent with increasing age of the host.The developing stages of S. angulata are found in the contorted tubules of the trunk-kidney but never invade the parenchymal tissue of the trunk or head-kidneys. The pansporoblasts developing in the wide proximal segment of the renal tubules, and the spores arising from the pansporoblasts are typically coelozoic parasites, for they apparently do not damage the tubular lining epithelium, nor its brush border. The main adverse effect of S. angulata is probably depression of resistance owing partly to deprivation of the host of nutrients secreted by the glomeruli and partly to mechanical obstruction of the distal tubular segments.

Notizen: Abstract. Viral haemorrhagic septicaemia (VHS) is reported for the first time in sea-water cultured rainbow trout. Heavy mortalities with typical signs and lesions A VHS virus (serotype 1) was isolated from the diseased fish. The mortalities were caused only by the VHS virus and 80 days post transfer of trout to sea-water the mortalities reached 85%, of the initial population.The disease was experimentally transmitted to rainbow trout, both in sea-water 3·104 pfu/ml of virus or by intramuscular injection of various doses of VHS1 (7·101 7·104 or 7·104 pfu per fish). Death occurred in all infected groups and started earlier in sea-water. Typical signs of VHS were observed in moribund fish. Viral multiplication was demonstrated to have occurred in fish organs.

Notizen: Abstract. Blood spots in the eye and haemorrhages in the musculature of the caudal peduncle were the main indication of disease in Australian hatchery reared brook trout Salvelinus fontinalis (Mitchill) and Atlantic salmon Salmo salar L. Mortalities of 0·1-0·5% per day occurred for 3–4 weeks in fish stocked in circular ponds in the months of February and March when temperatures were falling from 23–11·C. The bacterium belonging to the Family Enterobacteriaceae had similarities with Yersinia ruckeri (the RM bacterium) and Serratia liquefaciens. As a temporary designation SBS was used. It is recommended that its taxonomic position remain sub judice for the present. Inoculation experiments on brook trout and landlocked Atlantic salmon indicated that signs appeared 2–7 days after inoculation and that rainbow trout Salmo gairdneri Richardson were resistant to the disease.

Notizen: Abstract. Giemsa stained blood smears prepared from blennies caught on the shores of southern England and from cod taken from the Celtic and North seas revealed fish with clinical ‘piscine erythrocytic necrosis’ (PEN). The findings recorded here extend the distribution data on PEN infections and report that the infection in the blenny and cod may possibly have a similar developmental pattern. Data are presented on the prevalence of the infection in the fish population samples and on the proportion of infected cells in the smears examined.

Notizen: Books reviewed in this article:Tilapia—A guide to their biology and culture in Africa. By J. D. Balarin, assisted by J. P. Hatton.Pathophysiology—Altered regulatory mechanisms in disease 2nd ed. Ed. by E. D. Fkohuch.

Notizen: Abstract. A histopathological study was carried out on rainbow trout suffering from acute and chronic diplostomiasis. Clinical signs were few in acutely infected fish but in chronic cases fish were blind, exophthalmic and emaciated. Acute infections resulted in subcapsular cataract formation with varying capsular change. Chronically infected fish frequently showed lens dislocation, capsular rupture or duplication and the formation of Elschnig's pearls and Wedl cells. Phacogenic uveitis was seen in many fish with capsular rupture and retinal detachment frequently occurred as a sequal to vitreous detachment.

Notizen: Abstract. The intramolluscan development of Bunodera luciopercae was studied in laboratory infected pisidia maintained at three thermal regimes and compared with the seasonal development within a population of naturally infected first intermediate hosts at Llyn Tegid, Wales. At 4–8°C B. luciopercae did not develop beyond the sporocyst stage, although the development at 16–21°C We s similar to that reported by previous workers. The early stages of the intramolluscan development of B. luciopercae at Llyn Tegid were similar to the observed development at 16–21°C. However, the results suggest that falling autumn-winter temperatures may initiate the production of immature cercariae within first generation rediae. Cercariae matured and were shed at Llyn Tegid during a short period between March and May. Live, juvenile B. luciopercae were artificially transferred from recently killed perch Perca fluviatilis, gill netted from Llyn Tegid, into live uninfected, laboratory maintained perch. The effect of temperature on the development of B. luciopercae in its definitive host was investigated at four thermal regimes. At 16–20°C development did not proceed normally and the infection had disappeared after 130 days. At 4–8°C and at natural environmental temperatures, gravid flukes were produced after 158–168 days. An increase in temperature on infected fish maintained at 4–8°C for 43 days did not stimulate parasite maturation. It is suggested that B. luciopercae may require an extended period of vernalization to elicit maturation in its definitive host and that temperature has a major influence on the life history of this parasite.

Notizen: Abstract. Tetramicra brevifilum gen, n. sp, n. is described from the connective tissues throughout the body musculature of turbot Scophthalmus maximus (L.). Uninucleate spores (4.8·2 μm) are characterized by a large posterior vacuole, electron dense inclusion bodies in cytoplasm and nucleus and short filaments with 3–5 coils (50 μm extruded). Sporogony is apansporoblastic and tetrasporoblastic, within a host vacuole. Features of the xenoma (2 mm diameter) include a microvillous surface layer with microtubules and microfilaments and a reticulate nucleus with numerous nucleoli. Adherence between adjacent xenomas results in composite cysts of various sizes and shapes depending on site, being dendritic in the myomeres, arms extending between the fibres. Effects on the host, other than eventual destruction of the host cell, include displacement of muscle fibres in myomeres with loss of attachment to myocommata. Breakdown of the xenoma with release of spores into the connective tissue matrix was associated with a localized necrosis affecting surrounding muscle fibres, with leucocyte infiltration and collagen deposition. Spores phagocytosed by macrophages were destroyed in vacuoles, some with multi-laminate walls. Experimental infections established by intramuscular injection at marked sites remained localized, which suggested that xenomas afford sufficient scope for spore production without the need of secondary invasion. The swimming efficiency of turbot would be impaired in severe infections leading to lowered growth rates and increased mortality in wild populations due to predation and starvation. Feeding hierarchies exist in turbot and the disease could prove significant in farming conditions.

Notizen: Abstract. Since 1977, repeated outbreaks of infestation by Amyloodinium ocellatum (Dinoflagellida) in cultured gilthead bream Sparus aurata L. and seabass Dicen-trarchus labrax (L.) resulted in sporadic and mass mortalities. Fingerlings and yearlings as well as breeders were affected and, since 1978, larval and post-larval S. aurata in the hatchery. Infestations in juvenile and breeder fish were located predominantly on the gills and mucosal integument but in larval fish only the skin rather than the gills was infested. Histopathological changes of the infested gills are described. Vacuolar degeneration of the cells penetrated by the parasites’ rhizoids was evident at the site of attachment and cloudy swelling was apparent in the surrounding epithelial cells. In heavy infestations the entire epithelium of the gill filaments underwent hyperplasia with consequent fusion of the lamellae. Where hyperplasia was extensive the cells gradually degenerated while the inner epithelial layer showed distinct spongiosis and in some cases completely disintegrated. All heavily infested gills lacked mucous cells.

Notizen: Abstract. Virus was isolated from the brain and pancreatic tissues of Atlantic menhaden with ‘spinning’ disease. The biological, physical and chemical properties of the isolate were similar to those of infectious pancreatic necrosis virus. Inoculation of the virus into healthy menhaden resulted in the characteristic signs and mortality associated with the natural disease.