Animal Biowarfare Research: Historical Perspective and Potential Future Attacks

A biological attack on livestock or poultry could result in the loss of valuable animals, costs related to the containment of outbreaks and the disposal of carcasses, lost trade and other economic effects involving suppliers, transporters, distributors and restaurants; however, it is not possible to secure all livestock, and livestock are much less well guarded than human targets. Thus, the vulnerability of the livestock industry to the introduction of biological agents varies for the following reasons: (i) the majority of lethal and contagious biological agents are environmentally resilient, endemic in foreign countries and harmless to humans, making it easier for terrorists to acquire, handle and deploy these pathogens, (ii) with animals concentrated in fewer production facilities and frequently transported between these facilities, a single pathogen introduction could cause widespread infection and (iii) the extent of human travel around the globe makes it difficult to exclude exotic animal diseases as possible biological agents. Historically, many governments have developed and planned to use biological agents for direct attacks on livestock or poultry. In the past, developed nations have actively developed biological weapons to target animals. The potential spectrum of bioterrorism ranges from isolated acts against individuals by individuals to tactical and strategic military attacks and state‐sponsored international terrorism intended to cause mass casualties in animals, humans or both. This review provides an overview of the past development and use of biological weapons and describes potential future attacks.     

Evaluation of canine gastric motility with ultrasonography

For the evaluation of canine gastric motility with ultrasonography, contraction number of pyloric antrum and gastric emptying time (GET) by area and volume method developed by Bolondi et al.'s method were studied in 14 dogs. All experimental dogs were administered with saline and soup solution (10 ml/kg, B.W.). The mean values of contraction number of pyloric antrum in saline and soup group were 4.19 +/- 1.30/min and 4.82 +/- 0.65/min before feeding, and overall mean values were 4.66 +/- 1.37/min and 5.13 +/- 1.71/min, respectively. The mean values of the GET by area and volume method were 36.73 +/- 11.27, 40.00 +/- 8.87 min in saline group and 61.35 +/- 17.58, 59.11 +/- 14.46 min in soup group. In the GET in saline and soup groups, there was no significant difference between the area and volume method (p>0.05). Therefore, Bolondi et al.'s method by ultrasound can be used to evaluate the antropyloric motility and gastric emptying time with area and volume methods. The area method is easier to determine the GET than the volume method, but the latter is more accurate.     

A DNA Probe for Rapid Screening of Sugar Beet (Beta vulgaris L.) Carrying Extra Chromosomes from Wild Beets of the Procumbentes Section

Genes from wild species of the Procumbentes section can be transferred to sugar beet chromosomes via translocations. Since large translocations, including for example a gene for nematode resistance, generally result in meiotic disturbances, there is a need to select new diploid resistant beets from progenies of monosomic wild beet addition lines. A dispersed repetitive DNA probe, which is closely correlated with the resistance gene and hybridizes exclusively with wild beet DNA, appears to be highly superior to selection based on isozyme markers. Characteristic ‘fingerprints’ on the available monosomic addition lines reveal the existence of at last 5 different chromosomes in the wild species each housing a gene for nematode resistance. This probe can be used advantageously to identify individuals carrying the intact or fragmented wild beet chromosomes, or even various amounts of translocated chromatin. Strategies are discussed for the identification of new translocation types using straightforward squash dot or Southern hybridization techniques in combination with the wild beet DNA probe.     

Soundscapes reveal disturbance impacts: biophonic response to wildfire in the Sonoran Desert Sky Islands

Context

While remote sensing imagery is effective for quantifying land cover changes across large areas, its utility for directly assessing the response of animals to disturbance is limited. Soundscapes approaches—the recording and analysis of sounds in a landscape—could address this shortcoming.

Objectives

In 2011, a massive wildfire named “the Horseshoe 2 Burn” occurred in the Chiricahua National Monument, Arizona, USA. We evaluated the impact of this wildfire on acoustic activity of animal communities.

Methods

In 2013, soundscape recordings were collected over 9 months in 12 burned and 12 non-burned sites in four ecological systems. The seasonal and diel biological acoustic activity were described using the “Bioacoustic Index”, a detailed aural analysis of sound sources, and a new tool called “Sonic Timelapse Builder” (STLB).

Results

Seasonal biophony phenology showed a diurnal peak in June and a nocturnal peak in October in all ecological systems. On June mornings, acoustic activity was lower at burned than at non-burned sites in three of four ecological systems, due to a decreased abundance of cicadas directly impacted by the death of trees. Aural analyses revealed that 55% of recordings from non-burned sites contained insect sounds compared to 18% from burned sites. On October nights, orthopteran activity was more prevalent at some burned sites, possibly due to post-fire emergence of herbaceous.

Conclusions

Soundscape approaches can help address long-term conservation issues involving the responses of animal communities to wildfire. Acoustic methods can serve as a valuable complement to remote sensing for disturbance-based landscape management.

     

Pathogenicity of fungi associated with ash dieback symptoms of one‐year‐old Fraxinus excelsior in Montenegro

In addition to Hymenoscyphus fraxineus, two fungi identified as Diaporthe eres aff. and Fusarium sambucinum aff. were also isolated from necrotic bark lesions on declining one‐year‐old Fraxinus excelsior in a forest stand in Montenegro. To examine their involvement in ash decline, a pathogenicity test was performed using under bark inoculations on one‐year‐old Fraxinus excelsior. Hymenoscyphus fraxineus was included as comparison. All three fungal species proved highly pathogenic towards one‐year‐old seedlings although lesion sizes differed significantly between the different species. Hymenoscyphus fraxineus was most aggressive, followed by F. sambucinum aff., while D. eres aff. caused the smallest lesions. This study demonstrates for the first time the ability of isolates in the D. eres and F. sambucinum species complexes to cause decline on one‐year‐old common ash seedlings.     

Development of a joint system using a compressed wooden fastener I: evaluation of pull-out and rotation performance for a column—sill joint

Compressed wooden plates and dowels were used to connect members in post-and-beam structures as a substitute for a steel fastener. In order to take advantage of the characteristic properties of compressed wood and to achieve optimum joint performance, two compressed wooden plates were used in each joint to give multiple shear planes for each compressed wooden dowel. Consequently, this type of joint showed very good properties in pull-out and momen-trotation performance, and its engineering design could be further optimized. This joint is expected to be introduced to many kinds of structural systems, including long-span frame structures made of domestic timber found in Japanese residential houses.     

Characterization and application of recombinant β-glucosidase (BglH) from <Emphasis Type="Italic">Bacillus licheniformis</Emphasis> KCTC 1918

β-Glucosidase (β-1,4-D-glucoside glucohydrolase: EC.3.2.1.21) catalyzes the hydrolysis of β-glucosidic bonds between saccharides and aryl or alkyl groups. A gene encoding β-glucosidase from Bacillus licheniformis KCTC 1918, an anaerobic spore-forming soil bacterium, was cloned and characterized. The structural gene for the β-glucosidase consists of 1410 bp encoding 469 amino acid residues, and has a molecular weight of 53.4 kDa as estimated by sodium dodecyl sulfate polyacrylamide gel electrophoresis with 12% separating gel. The enzyme activity was determined against pNPG as a substrate. The enzyme was optimally active at pH 6.0 (citrate-phosphate buffer) and 47°C. β-Glucosidase retained 100% of its original activity for 24 h. The activity of the enzyme was stimulated by glycerol and urea and was decreased by Ca²⁺, Cu²⁺, Hg²⁺, Mg²⁺, and Mn²⁺. In particular, Cu²⁺ had the strongest negative effect on β-glucosidase activity. The purified β-glucosidase was active against pNPG and cellobiose. When the β-glucosidase was tested for cellulose hydrolysis, the supplement of β-glucosidase with cellulose increased the glucose yield from pine wood powder by 139.8%.     

Study on half-scale model test for light-frame shear wall

A half-scale model of a light-frame shear wall was developed to evaluate the racking performance of a fullscale shear wall (prototype). The effect of nail size on the performance of the shear wall was also investigated using models constructed with three types of nail. Materials for the model were determined through experimental methods, which included nail-head push-through, stud-to-sheathing nail connection, and static bending tests. Materials with which the model was made to be in similarity to the prototype were three-layer 4.8-mm plywood, 39.72-mm long nails, and 1 × 2 lumber cut from 2 × 4 studs. In accordance with ASTM E 72 and ASTM E 564, racking resistance tests were conducted on 20 shear walls. The results showed that the maximum load capacities of the prototype walls could be evaluated by the model without significantly different failure modes. Tests on the effect of nail size revealed that increasing the nail head diameter may improve the performance of shear walls.Parts of this paper were presented at the International Conference on Effective Utilization of Plantation Timber (ICEUPT'99), Chi-Tou, Taiwan, May 1999; and the World Conference on Timber Engineering (WCTE2000), Whistler, Canada, July–August 2000     

First report of Hymenoscyphus fraxineus on Fraxinus excelsior in Montenegro

During the monitoring of the mycological complex on different forest tree species in the Biogradska Gora National Park in north‐east Montenegro, symptoms indicative of ash dieback caused by Hymenoscyphus fraxineus were observed on young Fraxinus excelsior trees in the protected virgin forest, including dieback of plants and branches, wilting of leaves and shoots leading to a “flag‐like” habitus, premature shedding of leaves and longitudinal bark necroses. Using standard isolation methods, slow‐growing cultures with numerous phialides, typical of the asexual phase of the ash dieback fungus, were obtained. In addition, petioles with numerous characteristic apothecia were also recorded. This is the first report of H. fraxineus on common ash in Montenegro. Possible pathways of introduction and implications of the findings are discussed.     

Occurrence and pathogenicity of Phytophthora × cambivora on Prunus laurocerasus in Serbia

Cherry laurel (Prunus laurocerasus) is a native plant species in Serbian forests, but is also widely used for ornamental plantings. Following two extremely wet summers in 2014 and 2015, in spring and summer of 2016 and 2017, numerous cherry laurel plants with symptoms indicative for Phytophthora diseases, like wilting and chlorosis of leaves, dieback and bleeding bark necroses, were recorded in a park in Belgrade and in two ornamental nurseries in central Serbia. From necrotic bark samples and rhizosphere soil, self‐sterile Phytophthora isolates with woolly colonies were obtained. Due to the production of ellipsoid and elongated, non‐papillate sporangia in water and of ornamented oogonia with two‐celled antheridia in mating tests with tester strains of both Phytophthora × cambivora and P. cryptogea, these isolates were identified as P. ×cambivora which was confirmed by ITS sequence analysis. Pathogenicity of P. ×cambivora from cherry laurel (PCCL) was tested by inoculating one‐year‐old seedlings of cherry laurel under the bark. P. ×cambivora from European beech (PCB), and isolates of P. cactorum (CAC), P. cryptogea (CRY), P. plurivora (PLU) and P. ×serendipita (SER) were included as comparison. Three and a half months after inoculation, nine of the twelve plants in PCB, three in PCCL and CAC and two in PLU declined with longitudinal necroses and chlorosis, wilting and premature shedding of leaves. These results demonstrate the ability of P. ×cambivora to infect and cause decline of cherry laurel plants. The particularly high aggressiveness of the P. ×cambivora isolate from beech shows that this pathogen poses a serious risk to cherry laurel in the rare natural communities of cherry laurel and beech in Serbia.