Tusk abnormalities in wild boar (<Emphasis Type="Italic">Sus scrofa</Emphasis> L.)

This paper describes pathological alterations in the permanent canines (tusks) of four male wild boars. The mandibular tusks of all individuals, and also some of the maxillary tusks, exhibited an extended enamel hypoplasia in their apical portion, denoting an impairment of secretory ameloblast function. Moreover, the pulp cavities of the mandibular tusks were exposed through cleft-like openings in the wear (whetting) surfaces of the teeth. Presence of a plug of reparative dentine within the pulp cavity was observed in a split mandibular tusk of one individual. In a second boar, the presence of a plug of reparative dentine within the pulp cavity of the mandibular tusks was indicated radiographically. These findings suggested a reparation process attempting to demarcate a vital, apical pulp portion from a necrotic, incisal portion. The enamel hypoplasias observed in the teeth are regarded to be sequelae of the pulp inflammation caused by bacterial invasion in the mandibular tusks. Most likely, bacterial invasion of the dental pulp occurred through the cleft-like openings in the tusks whetting surfaces, the openings resulting from insufficient formation of secondary dentine. It is, however, also conceivable that pulp inflammation and partial necrosis occurred as a consequence of bacterial invasion of patent dentinal tubules, and that the openings in the whetting surface developed secondarily as a consequence of the pulp changes. One mandibular tusk showed marked signs of resorption apically, suggesting a spread of the inflammation from the pulp into the periodontium.     

Spleen size variations and reproduction in a Mediterranean population of wild boar (<Emphasis Type="Italic">Sus scrofa</Emphasis>)

We analysed spleen size variations of free-ranging wild boars from the west-central Iberian Peninsula during the hunting season (autumn and winter) in relation to the rut, the gestation effort and the attainment of sexual maturity by males and females. Females had larger spleens than males once they reached their sexual maturity. Individuals shot in winter had larger spleens than those shot in autumn, the start of the rutting period. In contrast to other reports, we found no influence of the reproductive status of adult females on their spleen sizes. Our findings may point to an influence of sex hormones, and possibly also stress hormones, and environmental factors on spleen development, mainly in adult and subadult males.     

Duplication polymorphism at MHC class II DRB1 locus in the wild boar (<Emphasis Type="Italic">Sus scrofa</Emphasis>)

The origin of allelic polymorphism in genes of the major histocompatibility complex represents a central topic in evolutionary genetics as it is probably the most polymorphic region in the nuclear genome of vertebrates. Accordingly, the analyses of genetic variability at these loci provide evidence complementary to the population genetics studies based on neutral loci. In this study, four wild boar populations, two from Italy (Florence region and Castelporziano Presidential Reserve, outside Rome) and one each from Hungary and Poland, were characterized at a highly polymorphic fragment including part of intron 1 and exon 2 of swine leukocyte antigen (SLA) class II DRB1 gene by direct sequencing and by cloning. Excluding the false alleles, a total of 18 different sequences were observed in 57 individuals. The high ratio of nonsynonymous (dN) vs synonymous (dS) substitution rates in the peptide-binding region supports the hypothesis that balancing selection is operating at this locus. A duplication event at the DRB1 gene was documented only in one Italian population with both copies being putatively active. This is the first evidence of a polymorphism for the number of copies of an SLA gene.     

Leptospira antibodies in wild boars (<Emphasis Type="Italic">Sus scrofa</Emphasis>) in Slovenia

Serum samples collected from 437 shot wild boars (Sus scrofa) were tested for the presence of antibodies against Leptospira interrogans sensu lato in wild boar in Slovenia. Assessment of leptospira-specific antibodies was performed by microscopic agglutination test. Antibodies against at least one of the pathogenic serovars were detected in 200 (45.8%) sera. From 200 positive samples, 100 samples (50%) had positive titre against a single serovar, while 100 (50%) samples had positive titres against two or more serovars. The most frequently detected antibodies were those against serovar Tarassovi. This investigation confirmed the presence of different pathogenic serovars in wild boar across Slovenia. It can be concluded that wild boars are natural reservoirs of at least some of the leptospiral serovars that represent a potential source of leptospirosis for other wild and domestic animals, as well as for humans.     

Habitat use and spatial patterns of wild boar <Emphasis Type="Italic">Sus scrofa</Emphasis> (L.): agricultural fields and edges

Rapidly increasing populations of wild boar in Sweden and Europe cause much damage to crops, and there is a critical need for more knowledge about their habitat utilization, especially of agricultural fields. In our study, we first assess the spatial pattern of damage in relation to the edges of agricultural fields. Next, with the aid of global positioning system collars, we studied the pattern of movement of wild boar on agricultural fields. Finally, in order to understand the role of agricultural fields, we studied how habitat selection may vary throughout the year. We found edge effects on damage patterns in agricultural fields. During winter and spring, we found wild boar not only to follow edges, but also to move along narrow landscape elements within agricultural fields. In our habitat analysis, we found strong avoidance of exposed agricultural fields throughout the year, but significantly less when crops are ripe.     

Reproductive parameters of wild boar (<Emphasis Type="Italic">Sus scrofa</Emphasis>) in three different parts of Germany

Recent increases in wild boar populations in central Europe necessitated an evaluation of the current reproductive performance of this species. During a 2-year study, samples of ovaries and uteri were taken from wild boars from areas in the (northern) state of Lower Saxony and the (more southern) Rhineland-Palatinate, Germany, in which wild boar have been vaccinated against classical swine fever (CSF) after several outbreaks of the disease. Serum samples were also analyzed to determine the prevalence of diseases that may influence pig reproduction and fertility. While there was a striking seasonality of reproductive performance, especially among older animals, mating was delayed in up to 15% of piglets and yearlings, leading to a second peak of farrowing from June to August. Depending on the area, between 60 and 70% of the piglets were most likely to become pregnant during the main period of reproduction, while another two thirds of the remaining individuals farrowed by the summer, which was the case in the mountainous areas. The arithmetic mean number of fetuses were 6.29 per piglet, 6.67 per yearling, and 7.64 per adult for wild boar in Eastern Lower Saxony; 4.58 per piglet and 4.63 per yearling and 6.56 per adult for wild boar in the Western Eifel and 4.77 per yearling and 6.50 per adult in the Palatine Forest, as the number of pregnant piglets has been too low to calculate an arithmetic mean in this area. The numbers of Lower Saxony wild boar fetuses per individual exceed previously known values. The results indicate that exogenous factors have a strong impact on both reproductive seasonality and the percentage of reproducing individuals in an age group. Ovulation rates, numbers of fetuses, and prevalence of pregnancies were found to be high among all age groups, while early onset of puberty and high pregnancy rates were typical of young animals (yearlings). The influence of some important reproductive diseases like Aujeszky’s disease, Brucellosis, porcine reproductive and respiratory syndrome, porcine parvovirosis, and CSF on wild boar reproduction and fertility was tested and found to be at least of minor interest for the reproductive outcome of the species.

Combined evaluation of bovine tuberculosis in wild boar (<Emphasis Type="Italic">Sus scrofa</Emphasis>) and red deer (<Emphasis Type="Italic">Cervus elaphus</Emphasis>) from Central-East Portugal

Bovine tuberculosis (bTB), widely considered a disease of livestock, can also affect large game species despite their poorly understood role in the epidemiology of the endemic state of the disease. The Idanha-a-Nova County, situated in the eastern region of central Portugal, is one of the most important regions for large game hunting activity. In order to investigate the role of large game and the presence of bTB in Idanha-a-Nova, 132 wild boar (Sus scrofa) and 339 red deer (Cervus elaphus) were inspected for bTB-compatible lesions (bTBCL) during the 2008–2009 hunting season. Tissue samples with bTBCL were collected and processed for bacteriological confirmation and spoligotyping of Mycobacterium bovis isolates. In wild boar, bTB lesions were single, mostly calcified (21/29, 72.4%) and situated in the mandibular lymph nodes. Lesions were more severe and largely non-calcified (36/40, 90%) in red deer. Wild boar presented a consistently higher bTB infection rate than the red deer; however, the difference was only significant in the southern B area. This area was significantly affected by bTB for both animal species (up to 72.7% in wild boar and 34.8% in red deer) and requires the urgent implementation of intervention measures to contain the disease. Wild boar may assume an important role in the emergence of new disease foci in distant areas that infected red deer cannot reach. Spoligotyping of M. bovis isolates grouped the strains into three clusters, revealing patterns common to wild boar, red deer, and also to local cattle, indicating possible transmission among different animal species.     

Population genetic structure of wild boar <Emphasis Type="Italic">Sus scrofa</Emphasis> in Bulgaria as revealed by microsatellite analysis

Over the last two decades wild boar Sus scrofa Linnaeus, 1758 became the most intensively managed game species in Bulgaria. In order to delineate the population genetic structure, which is essential for sustainable wildlife management, we screened 10 porcine microsatellites across 289 wild boar samples originating from all relevant bioregions of the country. Results based on F _ST values, Bayesian clustering methods and a multi-dimensional scaling analysis can be summarised as follows: (1) two main genetic groups were revealed for the Bulgarian data set: the first one included individuals collected from the Balkan Range Mountain and the northern part of the country and the second one comprised individuals from the Rhodope, Osogovo, Iskur Range and Rila mountains in southern and south-western Bulgaria; (2) all Bulgarian wild boar populations showed a higher level of genetic diversity compared to four populations from Germany which were included for outgroup comparison, and (3) wild boar sampled from a game enclosure were found to be genetically divergent from the other Bulgarian populations, indicating human impact on population genetic structure most likely resulted from fencing and former translocation actions. The evolutionary background leading to the two defined management units as well as conservation and management strategies are discussed.     

Mitochondrial and nuclear DNA analyses reveal pronounced genetic structuring in Tunisian wild boar <Emphasis Type="Italic">Sus scrofa</Emphasis>

We analysed 74 wild boars from Tunisia with respect to patterns of genetic differentiation and diversity based on sequences of the mitochondrial control region and genotypes at eight nuclear microsatellite loci. Analysis of molecular variance for both marker systems and Bayesian structure analysis of our microsatellite data revealed a clear break between northern and southern populations. Southern wild boar were monomorphic for one of three mtDNA haplotypes; the other two (one of which only occurred in three individuals) were confined to the north. A comparison with published sequences showed all three haplotypes to belong to the major European clade E1. Microsatellite diversity was similar to that found in earlier studies of wild boar (expected heterozygosity of 0.695 and 0.597 for the north and south, respectively). Contrary to the mtDNA results, we did not find unequivocal evidence of a bottleneck in Tunisian wild boar based on our microsatellite data. The clear distinction between northern and southern populations may be due to an Algerian origin of the southern animals.     

Dental anomalies in wild and domestic<Emphasis Type="Italic">Sus scrofa</Emphasis> in Illinois

The cleaned skulls of 39 wild and 30 domestic pigs from southern Illinois (USA) were assessed for dental anomalies including polydonty, oligodonty, misalignment, and rotation. Dental anomalies occurred in 16 wild and 15 domestic pigs. Oligodonty (either bilateral or unilateral) was the most common anomaly, occurring in 9 wild (23.1% of the sample) and 15 (50%) domestic pigs. In 22 of the 24 individuals exhibiting oligodonty, this anomaly involved the lower first premolar (P₁). Given the placement of P₁, oligodonty may reflect a trend toward reduction of the dental arcade from the primitive eutherian number. Domesticated species are reported to have more anomalies than wild counterparts because of inbreeding. We found no difference in the number of dental anomalies between domestic and wild pigs.     

Efficiency of spreading maize in the garrigues to reduce wild boar (<Emphasis Type="Italic">Sus scrofa</Emphasis>) damage to Mediterranean vineyards

The aim of this work was to assess the efficiency of dissuasive spreading of maize to reduce the level of wild boar damage to vines in a Mediterranean area (Puéchabon, southern France). The 50 wine growers of Puéchabon were all questioned about the annual losses caused by the wild boar in each vineyard of the study area between 1990 and 1992. We also studied the distribution of the damage on a smaller scale, by mapping the damaged vines within two vineyards. In summer 1993, we distributed 4.7 tons of maize in the woods , and then questioned the wine growers about the losses in each vineyard for 1993. During 1990–1992, on both large and small scales, the damage had a patchy distribution, with more patches of damage occurring close to the woods. Moreover, the later the vines ripened, the less frequent the damage. The severity of the damage was only affected by the distance of the vineyard from woods. In total, the wild boar consumed 20,049 kg of grapes each year between 1990–1992 (193 kg/ha), and 63% of the vineyards were damaged. In 1993, both the density and the compensation paid in the département increased threefold. However, in our study area, both the proportion of damaged vineyards (36%) and the level of damage in the vineyards (151 kg/ha) were reduced, saving more than 60% in compensation. The dissuasive spreading of maize is therefore an efficient tool to reduce the level of damage to vineyards.     

Environmental correlates of nest site selection by wild boar<Emphasis Type="Italic">Sus scrofa</Emphasis>

Location of nests, built by free-living wild boarsSus scrofa Linnaeus, 1758, was analysed in order to assess the importance of environmental and social conditions in piglets’ survival during the first days after birth. The research was carried in a Mediterranean forest with different habitats and high density of natural predators. The results indicate that most nests were already constructed before the beginning of March, being located preferentially in areas with: (1) abundant plant cover, (2) water nearby and (3) a warmer temperature than in other places. These results suggest that female wild boar try to diminish mortality caused by natural predators by constructing their nests in places with dense cover. Water is very important because of the females’ new necessities derived from milk production. Warm nesting places might diminish deaths after birth in a species with important thermoregulation deficiencies. The distance between nests is also important because in this period the wild boar develops a territorial behaviour, possibly with the objective of establishing bonds between the mother and her offspring before returning to the familiar units made up of several females and their piglets, all of them of a similar age.     

Fine mapping of a male sterility gene <Emphasis Type="Italic">ms</Emphasis>-<Emphasis Type="Italic">3</Emphasis> in a novel cucumber (<Emphasis Type="Italic">Cucumis sativus</Emphasis> L.) mutant

Key message

The cucumber male sterility gene ms - 3 was fine mapped in a 76 kb region harboring an MMD1 -like gene Csa3M006660 that may be responsible for the male sterile in cucumber.

Abstract

A cucumber (Cucumis sativus L.) male sterile mutant (ms-3) in an advanced-generation inbred line was identified, and genetic analysis revealed that the male sterility trait was controlled by a recessive nuclear gene, ms-3, which was stably inherited. Histological studies suggested that the main cause of the male sterility was defective microsporogenesis, resulting in no tetrad or microspores being formed. Bulked segregant analysis (BSA) and genotyping of an F₂ population of 2553 individuals were employed used to fine map ms-3, which was delimited to a 76 Kb region. In this region, a single non-synonymous SNP was found in the Csa3M006660 gene locus, which was predicted to result in an amino acid change. Quantitative RT-PCR analysis of Csa3M006660 was consistent with the fact that it plays a role in the early development of cucumber pollen. The protein encoded by Csa3M006660 is predicted to be homeodomain (PHD) finger protein, and the high degree of sequence conservation with homologs from a range of plant species further suggested the importance of the ms-3 non-synonymous mutation. The data presented here provide support for Csa3M006660 as the most likely candidate gene for Ms-3.

     

<Emphasis Type="Italic">Salmonella enterica</Emphasis> subspecies <Emphasis Type="Italic">enterica</Emphasis> serovar Choleraesuis in a wild boar population in Germany

Salmonella (S.) enterica subspecies enterica serovar Choleraesuis, the swine-adapted serovar is found rarely in Western European countries including Germany. However, the regional laboratory of the federal state Thuringia in Germany examined diseased wild boars routinely also for the occurrence of Salmonella organisms. Between 2006 and 2008, only the serovar S. Choleraesuis was islolated from 24 animals, three strains isolated from domestic pigs were included. In order to detect a possible epidemiological context, the strains of S. Choleraesuis were characterised by macrorestriction and plasmid analysis, repetitive sequence PCR, antimicrobial testing and determining the biochemical profile. A combination of all methods enabled the identification of five epidemiological groups. Two groups were detected in the same territory but three other discriminative groups were predominant in different regions. S. Choleraesuis strains of the different epidemiological groups circulate in wild boar populations in the corresponding regions. However, it could be concluded that both natural barriers like mountains and artificial barriers like arterial roads may cause the separation of wild boar populations and as a result also the respective S. Choleraesuis organisms. The occurrence of the identical epidemiological groups in wild boars and domestic pigs indicates the possible mutual exposure of the pathogen. To avoid risks for human and domestic pig health regular inspection of meat from wildlife by official veterinarians and advice of hunters and persons who prepare and consume wild boar meat should be enhanced.     

<Emphasis Type="Italic">Agrobacterium-</Emphasis>mediated transformation of gypsophila (<Emphasis Type="Italic">Gypsophila paniculata</Emphasis> L.)

As a major contributor to the flower market, Gypsophila paniculata is an important target for the breeding of new varieties. However, gypsophila breeding is strongly hampered by the sterility of this species’ genotypes and the lack of a genetic-transformation procedure for this genus. Here we describe the establishment of a transformation procedure for gypsophila (Gypsophila paniculata L.) based on Agrobacterium inoculation of highly regenerative stem segments. The transformation procedure employs stem explants derived from GA₃-pretreated mother plants and a two-step selection scheme. The GA₃ treatment was crucial for obtaining high gene-transfer frequencies (75–90% GUS-expressing explants out of total inoculated explants), as shown using three different gypsophila varieties. An overall transformation efficiency of five GUS-expressing shoots per 100 stem explants was demonstrated for cv. Arbel. The applicability of the transformation system to gypsophila was further reinforced by the generation of transgenic plants expressing Agrobacterium rhizogenes rolC driven by a CaMV 35S promoter. Transgenic gypsophila plantlets exhibited extensive rooting and branching, traits that could be beneficial to the ornamental industry.     

Simple sequence repeat analyses of interspecific hybrids and MAALs of <Emphasis Type="Italic">Oryza </Emphasis><Emphasis Type="Italic">officinalis</Emphasis> and <Emphasis Type="Italic">Oryza sativa</Emphasis>

Wild rice is a valuable resource for the genetic improvement of cultivated rice (Oryza sativa L., AA genome). Molecular markers are important tools for monitoring gene introgression from wild rice into cultivated rice. In this study, Simple sequence repeat (SSR) markers were used to analyze interspecific hybrids of O. sativa-O. officinalis (CC genome), the backcrossing progenies and the parent plants. Results showed that most of the SSR primers (335 out of 396, 84.6%) developed in cultivated rice successfully amplified products from DNA samples of wild rice O. officinalis. The polymorphism ratio of SSR bands between O. sativa and O. officinalis was as high as 93.9%, indicating differences between the two species with respect to SSRs. When the SSR markers were applied in the interspecific hybrids, only a portion of SSR primers amplified O. officinalis-specific bands in the F(1) hybrid (52.5%), BC(1) (52.5%), and MAALs (37.0%); a number of the bands disappeared. Of the 124 SSR loci that detected officinalis-specific bands in MAAL plants, 96 (77.4%) showed synteny between the A and C-genomes, and 20 (16.1%) showed duplication in the C-genome. Sequencing analysis revealed that indels, substitution and duplication contribute to the diversity of SSR loci between the genomes of O. sativa and O. officinalis.     

Factors affecting <Emphasis Type="Italic">Agrobacterium</Emphasis>-mediated transformation in <Emphasis Type="Italic">Hybanthus</Emphasis><Emphasis Type="Italic">enneaspermus</Emphasis> (L.) F. Muell.

Agrobacterium tumefaciens-mediated transformation system was established for Hybanthus enneaspermus using leaf explants with the strain LBA4404 harbouring pCAMBIA 2301 carrying the nptII and gusA genes. Sensitivity of leaf explants to kanamycin was standardized (100 mg/l) for screening the transgenic plants. Transformation parameters (OD, virulence inducer, infection time, co-cultivation period, bactericidal antibiotics, etc.) influencing the gene transfer and integration were assessed in the present investigation. Fourteen-day pre-cultured explants were subjected with Agrobacterium strain LBA4404. Optimized parameters such as culture density of 0.5 OD₆₀₀, infection time of 6 min, AS concentration of 150 µM with 3 days co-cultivation revealed maximum transformation efficiency based on GUS expression assay. The presence of gusA in transgenics was confirmed by polymerase chain reaction and Southern blotting analysis. The present transformation experiment yielded 20 shoots/explant with higher transformation efficiency (28 %). The protocol could be used to introduce genes for trait improvement as well as for altering metabolic pathway for secondary metabolites production.     

Genetics and fine mapping of a yellow-green leaf gene (<Emphasis Type="Italic">ygl</Emphasis>-<Emphasis Type="Italic">1</Emphasis>) in cabbage (<Emphasis Type="Italic">Brassica oleracea</Emphasis> var. <Emphasis Type="Italic">capitata</Emphasis> L.)

Cabbage (Brassica oleracea var. capitata L.) is one of the most popular cultivated vegetables worldwide. Cabbage has rich phenotypic diversity, including plant height, head shape, head color, leaf shape and leaf color. Leaf color plays an important role in cabbage growth and development. At present, there are few reports on fine mapping of leaf color mutants in B. oleracea. In this study, a naturally occurring yellow-green leaf cabbage mutant (YL-1), derived from the self-pollinated progenies of the hybrid ‘Hosom’, was used for inheritance analysis and gene mapping. Segregation populations including F₂ and BC₁ were generated from the cross of two inbred lines, YL-1 and 01–20. Genetic analysis with the F₂ and BC₁ populations demonstrated that the yellow-green leaf color was controlled by a single recessive nuclear gene, ygl-1. Insertion–deletion (InDel) markers, designed based on the parental re-sequencing data, were used for the preliminary mapping with BSA (bulked segregant analysis) method. A genetic map constructed with 15 InDels indicated that ygl-1 was located on chromosome C01. The ygl-1 gene is flanked by InDel markers ID2 and M8, with genetic distances of 0.4 cM and 0.35 cM, respectively. The interval distance between two markers is 167 kb. Thus, it enables us to locate the ygl-1 gene for the first time in B. oleracea. This study lays the foundation for candidate gene prediction and ygl-1gene cloning.     

<Emphasis Type="Italic">Ty</Emphasis>1<Emphasis Type="Italic">/copia</Emphasis>- and <Emphasis Type="Italic">Ty</Emphasis>3<Emphasis Type="Italic">/gypsy</Emphasis>-like DNA sequences in <Emphasis Type="Italic">Helianthus </Emphasis>species

Two repeated DNA sequences isolated from a partial genomic DNA library of Helianthus annuus, p HaS13 and p HaS211, were shown to represent portions of the int gene of a Ty3 /gypsy retroelement and of the RNase-Hgene of a Ty1 /copia retroelement, respectively. Southern blotting patterns obtained by hybridizing the two probes to BglII- or DraI-digested genomic DNA from different Helianthus species showed p HaS13 and p HaS211 were parts of dispersed repeats at least 8 and 7 kb in length, respectively, that were conserved in all species studied. Comparable hybridization patterns were obtained in all species with p HaS13. By contrast, the patterns obtained by hybridizing p HaS211 clearly differentiated annual species from perennials. The frequencies of p HaS13- and p HaS211-related sequences in different species were 4.3x10(4)-1.3x10(5) copies and 9.9x10(2)-8.1x10(3) copies per picogram of DNA, respectively. The frequency of p HaS13-related sequences varied widely within annual species, while no significant difference was observed among perennial species. Conversely, the frequency variation of p HaS211-related sequences was as large within annual species as within perennials. Sequences of both families were found to be dispersed along the length of all chromosomes in all species studied. However, Ty3 /gypsy-like sequences were localized preferentially at the centromeric regions, whereas Ty1/ copia-like sequences were less represented or absent around the centromeres and plentiful at the chromosome ends. These findings suggest that the two sequence families played a role in Helianthusgenome evolution and species divergence, evolved independently in the same genomic backgrounds and in annual or perennial species, and acquired different possible functions in the host genomes.