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.     

A serological survey of selected pathogens in wild boar (<Emphasis Type="Italic">Sus scrofa</Emphasis>) in northern Turkey

During the hunting season in March 2012, a total of 93 blood samples were collected from wild boars (Sus scrofa) shot in the area of northern Turkey (Samsun and Gumushane provinces). These blood samples were examined by enzyme immunoassay (ELISA) for the presence of antibodies to classical swine fever virus (CSFV), Aujeszky’s disease virus (ADV), porcine reproductive and respiratory syndrome virus (PRRSV), porcine respiratory coronavirus (PRCV), swine influenza virus (SIV), porcine parvovirus (PPV), swine vesicular disease virus (SVDV), hepatitis E virus (HEV), African swine fever virus (ASFV), porcine rotavirus (PRV), transmissible gastroenteritis virus (TGEV) and bovine viral diarrhoea virus (BVDV). Out of 93 serum samples examined, 65 (69.9 %) were positive for PRV, 22 (23.7 %) were positive for ADV, 5 (5.4 %) were positive for BVDV, 4 (4.3 %) were positive for PPV and 2 (2.2 %) were positive for PRRSV. All sera were negative for ASFV, SVDV, HEV, SIV, PRCV, TGEV and CSFV. The results, recorded for the first time in Turkey, supported the hypothesis that wild boar act as a potential reservoir of selected viruses and thus have a role in the epidemiology of these diseases.     

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.     

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.     

Response to human presence during nocturnal line transect surveys in fallow deer (<Emphasis Type="Italic">Dama dama</Emphasis>) and wild boar (<Emphasis Type="Italic">Sus scrofa</Emphasis>)

The use of line transect methodology and portable thermal imaging for ground survey of wildlife should require a good knowledge of the behavioural response of the animals to the presence of an observer, in order to take into account the potential bias in density estimate caused by deviation from the assumption that distances are recorded at the initial position. We used ten fallow deer and eight wild boar fitted with radiocollars to investigate animals’ response during simulated nocturnal line transect surveys, carried out in a Mediterranean plain forest. The experiment consisted in radiolocating a focal animal before and after an observer walked a transect nearby (<100 m). Each transect line was followed using a Global Positioning System (GPS) navigator. We carried out a total of 64 trials on fallow deer and 57 on wild boar. Results showed that despite most of the animals moved significantly in response to the observer (mean ± standard error, wild boar—95.3 ± 10.0 m; fallow deer—149.6 ± 14.2 m), the flying patterns were different in the two species: the reaction of fallow deer turned out to be more intense and directional with respect to that showed by wild boar. Although these results sound explorative, the experiment attempted here, for the first time, is relevant for an appropriate design of nocturnal distance-sampling surveys and gives information about potential bias arising from animal’s behavioural response. We believe that these first results may foster more in-deep analyses which are now made possible with the adoption of GPS technology for animal location.     

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.     

Ectopic expression of cucumber (<Emphasis Type="Italic">Cucumis sativus</Emphasis> L.) <Emphasis Type="Italic">CsTIR</Emphasis>/<Emphasis Type="Italic">AFB</Emphasis> genes enhance salt tolerance in transgenic <Emphasis Type="Italic">Arabidopsis</Emphasis>

Auxin receptors TIR1/AFBs play an essential role in a series of signaling network cascades. These F-box proteins have also been identified to participate in different stress responses via the auxin signaling pathway in Arabidopsis. Cucumber (Cucumis sativus L.) is one of the most important crops worldwide, which is also a model plant for research. In the study herein, two cucumber homologous auxin receptor F-box genes CsTIR and CsAFB were cloned and studied for the first time. The deduced amino acid sequences showed a 78% identity between CsTIR and AtTIR1 and 76% between CsAFB and AtAFB2. All these proteins share similar characteristics of an F-box domain near the N-terminus, and several Leucine-rich repeat regions in the middle. Arabidopsis plants ectopically overexpressing CsTIR or CsAFB were obtained and verified. Shorter primary roots and more lateral roots were found in these transgenic lines with auxin signaling amplified. Results showed that expression of CsTIR/AFB genes in Arabidopsis could lead to higher seeds germination rates and plant survival rates than wild-type under salt stress. The enhanced salt tolerance in transgenic plants is probably caused by maintaining root growth and controlling water loss in seedlings, and by stabilizing life-sustaining substances as well as accumulating endogenous osmoregulation substances. We proposed that CsTIR/AFB-involved auxin signal regulation might trigger auxin mediated stress adaptation response and enhance the plant salt stress resistance by osmoregulation.     

<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.     

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">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.     

<Emphasis Type="Italic">Agrobacterium</Emphasis>-mediated genetic transformation of grapevine (<Emphasis Type="Italic">Vitis vinifera</Emphasis> L.) with a novel stilbene synthase gene from Chinese wild <Emphasis Type="Italic">Vitis pseudoreticulata</Emphasis>

A novel stilbene synthase gene (STS), cloned from Chinese wild Vitis pseudoreticulata (W. T. Wang) and responsible for synthesis of the phytoalexin resveratrol in grapevine, was successfully transferred into V. vinifera L. cv. Thompson Seedless via Agrobacterium tumefaciens-mediated transformation. Using transformation procedures developed in the present study, 72% GFP-positive germinated embryos were produced with about 38% of transformed embryos regenerated into normal plantlets. Integration of the STS gene into the transgenic plants was verified by PCR and Southern blot analysis. Expression of the STS gene was detected by high performance liquid chromatography (HPLC), which showed that the resveratrol concentration in the transgenic plants was 5.5 times higher than that in non-transformed control plants. Chaohong Fan and Ni Pu contributed equally to this work.