Pollution related effects on immune function and stress in a free-living population of pied flycatcher Ficedula hypoleuca

We investigated whether exposure to heavy metal pollution affected the immune function of individuals in a free living population of a small insectivorous passerine bird, the pied flycatcher Ficedula hypoleuca. We measured humoral immune responses in two study areas: a polluted area in the vicinity of a copper smelter and a control area far from the smelter. Plasma corticosterone level and blood heterophil/lymphocyte ratio (H/L) were used as more general physiological measures of stress. The immune response of F hypoleuca was not suppressed by pollution stress. In contrast, we found that F hypoleuca males showed stronger Immoral immune responses to a novel antigen (tetanus toxoid) in the polluted environment than in the unpolluted one. After the immunization of males, numbers of lymphocytes rose significantly more in the polluted area, leading to a smaller H/L ratio than in males from the control area. Females showed no pollution related effects on their immune responses. Corticosterone levels of males and nestlings were not related to pollution levels. Nestlings showed somewhat higher H/L ratios and lower fledging success in the polluted area, both factors indicating increased stress levels in a polluted area. Our results suggest that Immoral immune response of male F hypoleuca may be enhanced under moderate levels of heavy metal Pollution. Enhanced immune function may, however, also be costly for birds and the higher humoral immune responses in polluted areas may thus have negative effects on the birds' breeding performance and survival

Experimental evidence for major histocompatibility complex-allele-specific resistance to a bacterial infection

The extreme polymorphism found at some major histocompatibility complex (MHC) loci is believed to be maintained by balancing selection caused by infectious pathogens. Experimental support for this is inconclusive. We have studied the interaction between certain MHC alleles and the bacterium Aeromonas salmonicida, which causes the severe disease furunculosis, in Atlantic salmon (Salmo salar L.). We designed full-sibling broods consisting of combinations of homozygote and heterozygote genotypes with respect to resistance or susceptibility alleles. The juveniles were experimentally infected with A. salmonicida and their individual survival was monitored. By comparing full siblings carrying different MHC genotypes the effects on survival due to other segregating genes were minimized. We show that a pathogen has the potential to cause very intense selection pressure on particular MHC alleles; the relative fitness difference between individuals carrying different MHC alleles was as high as 0.5. A co-dominant pattern of disease resistance/susceptibility was found, indicative of qualitative difference in the immune response between individuals carrying the high- and low-resistance alleles. Rather unexpectedly, survival was not higher among heterozygous individuals as compared with homozygous ones

Adaptive and Neutral Genetic Variation and Colonization History of Atlantic Salmon, Salmo salar

I combined neutral microsatellite markers with the major histocompatibility complex (MHC) class IIB to study genetic differentiation and colonization history in Atlantic salmon, Salmo salar, in the Baltic Sea and in the north-eastern Atlantic. Baltic salmon populations have lower levels of microsatellite genetic variation, in terms of heterozygosity and allelic richness than Atlantic populations, confirming earlier findings with other genetic markers, suggesting that the Baltic Sea populations have been exposed to genetic bottlenecks, most likely at a founding event. On the other hand, the level of MHC variation was similar in the Baltic and in the north-eastern Atlantic, indicating that positive balancing selection has increased the level of MHC-variation. Both microsatellite and MHC class IIB genetic variation give strong support to the hypothesis that the Baltic salmon are of a biphyletic origin, the southern population in this study is strongly differentiated from both the northern Baltic salmon populations and from the north-eastern Atlantic populations. Salmon may have colonized the northern Baltic Sea either from the south, via the so called “Närke strait” or from the north, via a proposed historical connection between the White Sea and the northern Baltic. At microsatellites, no significant isolation-by distance was found at either colonization route. At the MHC, populations were significantly isolated by distance when assuming that colonization occurred via the “Närke strait”

Genetic Variation in Mhc Class II B in Atlantic Salmon: Evolutionary and Ecological Perspectives

The Major Histocompatibility Complex (Mhc) genes are crucial for the recognition of self and non-self peptides by the immune defence system. They are extremely variable in most studied vertebrate species. I have studied the extent and the importance of Mhc variation in Atlantic salmon (Salmo salar), mainly from the Baltic Sea. Levels of Mhc heterozygosity in hatchery populations of salmon were between 0.5 and 0.8, the lowest values in a hatchery where a lower annual number of breeders were used. In a wild, naturally spawning population, heterozygosity was 0.7, which is in between the levels observed in the hatcheries. The origin and maintenance of the high level of Mhc variation is a contentious issue. My results show that there is only one single Mhc class II B gene in Atlantic salmon, and indicate that point mutations are not the sole origin of variation in this single gene. New alleles appear to result from recombination. Mhc variation is assumed to be maintained by some kind of balancing selection, which is supported by my results. Balancing selection may be in terms of over-dominance selection (heterozygote advantage) or frequency dependent selection. I have not found any support for over-dominance selection. I found that certain Mhc alleles were associated with furunculosis, a bacterial infection as well as the syndrome M74, which has caused high mortality in Baltic Sea salmon. There was however, no association between heterozygosity and either furunculosis or M74. These results, together with the lack of an excess of heterozygous individuals in the natural population, support that frequency dependent selection rather than over-dominant selection, is the main mechanism for maintaining Mhc variation

The Baltic salmon in battle with pollutants and disease. / Östersjölaxen i kamp mot miljögifter och sjukdom.

The Baltic salmon has during the latter part of the 20th century been subjected to several threats. Most notable have been the high levels of contaminant release and a disease that causes early life mortality of the young. Ever since utilization of water power from the Swedish rivers began in the 1940ies, thus deminishing the natural breeding areas for the salmons, an extensive hatchery program has compensated for the loss and, generally, increased the salmon populations in the Baltic Sea but simultanously decreased the genetic variation. In the 70ies a disease appeared in the hatcheries, causing high mortality (up to 95%) of the salmon fry. The explanation for this devastating syndrome, known today as M74, has not been found but it has been suggested that the high load of persistent environmental pollutants like PCB and dioxin that historically has been released into the water systems, have contributed to the problem. Since these chemicals strongly resemble endogenous signalling substances -hormones- exposed individuals experience altered cellular signalling which eventually results in cell death. This signalling is mediated by a receptor protein known as the aryl hydrocarbon (Ah) receptor which during recent years has been subjected to much interest because of its role in mediating toxicity of several environmental pollutants. Further research on the Ah receptor may in the future bring greater understanding of some the problems that the Baltic salmons presently are suffering from

Inter- and intralocus recombination drive MHC class IIB gene diversification in a teleost, the three-spined stickleback Gasterosteus aculeatus

The mutational mechanism underlying the striking diversity in MHC (major histocompatibility complex) genes in vertebrates is still controversial. In order to evaluate the role of inter- and intragenic recombination in MHC gene diversification, we examined patterns of nucleotide polymorphism across an exon/intron boundary in a sample of 31 MHC class IIB sequences of three-spined stickleback (Gasterosteus aculeatus). MHC class IIB genes of G. aculeatus were previously shown to be under diversifying (positive) selection in mate choice and pathogen selection experiments. Based on recoding of alignment gaps, complete intron 2 sequences were grouped into three clusters using maximum-parsimony analysis. Two of these groups had > 90% bootstrap support and were tentatively assigned single locus status. Intron nucleotide diversity within and among loci was low (p-distance within and among groups = 0.016 and 0.019, respectively) and fourfold lower than the rate of silent mutations in exon 2, suggesting that noncoding regions are homogenized by frequent interlocus recombination. A substitution analysis using GENECONV revealed as many intergenic conversion events as intragenic ones. Recombination between loci may explain the occurrence of sequence variants that are particularly divergent, as is the case in three-spined stickleback, with nucleotide diversity attaining d(N) = 0.39 (peptide-binding residues only). For both MHC class II loci we also estimated the amount of intragenic recombination as population rate (4N(e)r) under the coalescent and found it to be approximately three times higher compared to point mutations (Watterson estimate per gene, 4N(e)mu). Nonindependence of molecular evolution across loci and frequent recombination suggest that MHC class II genes of bony fish may follow different evolutionary dynamics than those of mammals. Our finding of widespread recombination suggests that phylogenies of MHC genes should not be based on coding segments but rather on noncoding introns

Allelic polymorphism in MHC class IIB in four populations of Atlantic salmon (Salmo salar)

We sequenced exon 2 of the MHC class II B gene in Atlantic salmon from the Baltic Sea and identified 17 different exon 2 alleles among 22 different restriction fragment length polymorphism haplotypes. The sequences differed at between I and 34 bases. Two different tests were used to estimate the importance of recombination in the generation of new alleles. Recombination events appear to have occurred between three and nine times. Only two pairs of sequences differed by less than five nucleotides', minimizing the importance of point mutations for generating new alleles. Phylogenetic analysis showed that sequences did not cluster according to populations, and genetic distances between populations were small compared to those obtained by allele frequency data. These results, together with the similarity found between exon 2 sequences from Baltic salmon and Norwegian salmon, indicate that all of the identified alleles were present in the ancient salmon population colonizing the Baltic rivers after the last glaciation

Extra-pair fertilizations in the Sedge Warbler

Parentage of 201 young (from 44 broods) in a population of Sedge Warblers Acrocephalus schoenobaenus in South Central Sweden, 1990-1992, was determined by using multilocus DNA fingerprinting. The frequency of extra-pair young (EPY) was 7.5% and they occurred in 23% of the broods. For 11 out of 15 extra-pair young we could identify the true father; in all of the cases he was a close neighbour. Among ten broods with extra-pair young, nine contained only one extra-pair sire and the remaining brood two extra-pair sires. The frequency of EPF varied among years (1.8-11.8%). The seasonal timing of broods with and without extra-pair young did not differ, and the occurrence of extra-pair fertilizations (EPF) was not related to the length of the pair male's mate-guarding period. EPF were not related to breeding synchrony (estimated as the mean number of fertile females per day during a female's fertile period). Pairs whose nests contained extra-pair young had more territories within 100 m of their nest than pairs without extra-pair young. Females that engaged in EPF had fewer attractive (i.e. singing) males to choose among the day before and at the day they formed their pair bend. Moreover, in all but one case the extra-pair male was not singing (i.e. not available as pair mate) the day the EPF-female settled. Males that fertilized extra-pair young tended to arrive earlier and to have higher pairing success than both males that were cuckolded and other males. Hence, female Sedge Warblers engaged in extra-pair fertilizations with attractive male neighbours. These results are consistent with the hypotheses that females participating in EPF are seeking genetic benefits to their offspring, but we cannot exclude the alternative explanation that attractive males are more efficient in forcing females to accept EPF

Allelic variation of Mhc class II in Atlantic salmon; a population genetic analysis

We have studied the degree of genetic variation at a variable Mhc class II beta gene in four populations of Atlantic salmon Salmo salar by using RFLPs. The class II beta gene encodes the part of the Mhc class II molecule that contains the antigen binding region and is therefore essential for disease resistance. There was extensive genetic variation in all four populations; the expected heterozygosity (H-si) varied between 0.50 and 0.81. Heterozygosity tended to be higher in broods surviving a syndrome causing high mortalities (60-95%) in Swedish salmon hatcheries. Populations that had experienced more incidences of genetic bottlenecks (years when fewer than 60 adults had been used for breeding) and had a lower average effective population size (N-e), had a lower degree of heterozygosity. The four populations differed significantly in allele frequencies as measured by F-ST = 0.13 and Nei's genetic distance (D = 0.09 - infinity). Pairwise F-ST values varied between 0.01 and 0.23, all but one being highly significant, indicating a differentiation between the populations in Mhc class II beta. This study shows that the four populations of Atlantic salmon have a high degree of polymorphism in the Mhc class II beta gene. However, there was great variation between different hatcheries, both in heterozygosity levels and allele frequencies