Genotype characterisation of Giardia duodenalis isolates from domestic and farm animals by SSU-rRNA gene sequencing

In order to investigate the genotypes of Giardia duodenalis from domestic and farm animals in Italy, 21 Giardia isolates, 17 from dogs, 1 from cat and 3 from dairy calves, were genetically characterised by SSU-rRNA gene sequencing. Among dogs, 76.5% of isolates showed the dog-specific genotypes (Assemblages C, D and C/D mixed Assemblage) and 23.5% exhibit potential zoonotic genotypes (Assemblage A and A/C mixed Assemblages). The cat isolate belonged to assemblage A, whereas the sequences among the isolates from calves were found to correspond to hoofed-livestock genotype, namely Assemblage E. These findings suggest that infection of humans by zoonotic genotypes from domestic animals could be of low epidemiological significance, although possible. The present study represents the first contribute to the knowledge of G. duodenalis genotypes in domestic and farm animals from Italy.     

Evaluating the distribution of priority benthic habitats through a remotely operated vehicle to support conservation measures off Linosa Island (Sicily Channel,Mediterranean Sea)

1. Explorations of the Mediterranean deep sea using remotely operated vehicles have shown that the sea bed hosts rich habitats, supporting high biodiversity. However, there have been only a few studies dealing with the southern part of the basin, leading to limited protection and conservation efforts in this area.
2. This study aimed to explore the sea bed off Linosa Island (Sicily Channel, southern Mediterranean Sea), which is considered a ‘sentinel area’ for alien species and global environmental changes owing to its geographic position, thus deserving special attention.
3. Remotely operated vehicle surveys, carried out in 2016 and 2017, were analysed to provide the first ecological characterization of benthic assemblages at depths −19 – −384 m around Linosa Island.
4. Communities were dominated by three priority habitats, amounting to 39% of the almost 5 km of the sea floor that was surveyed. These are represented in the euphotic zone by Posidonia oceanica meadows and, at greater depth, by newly discovered dense coral forests and extended rhodolith/maërl beds. Sixteen habitat-forming species included in the Red List of the International Union for Conservation of Nature (e.g. gorgonians Eunicella cavolini and Paramuricea clavata, and black corals Antipathella subpinnata and Leiopathes glaberrima) were recorded, as well as individuals of Sargassum sp. at −100 m depth.
5. The volcanic island of Linosa represents a small, naturally preserved area, with very limited human pressure, hosting rich marine benthic biodiversity. Given the high species and habitat richness, we recommend its inclusion in the Special Protected Areas of Mediterranean Importance project (United Nations Environment Programme) and suggest a redefinition of the existing marine protected area extension.

     

Fungal root pathogen (<Emphasis Type="Italic">Heterobasidion parviporum</Emphasis>) increases drought stress in Norway spruce stand at low elevation in the Alps

Tree-ring patterns of Picea abies (L.) Karst. both unaffected and affected by Heterobasidion parviporum were analysed in three mature stands located at different elevations in the Eastern Alps. The main objectives were (1) to clarify the role of climatic conditions on infected trees; (2) to estimate indirect volume losses due to the prolonged presence of the fungus within the wood. The low elevation site showed the highest growth decline in the last decade, whereas all infected trees at medium and high elevation showed a slow growth decline over many decades. We hypothesise that infection could be dated over 80 years at the highest site. Fungal attack made P. abies more susceptible to drought stress at low elevation site. Both infected and healthy P. abies at medium and high elevation showed similar climate–growth relationships, suggesting that the same driving environmental factors influence their growth. At low elevation, H. parviporum was seemingly more aggressive, causing a more rapid decline, decreasing the ability of host trees to cope with drought and, in some cases, inducing cambial activity to stop. P. abies at higher elevation, however, exhibited a very slow decline and no sign of increasing water stress since the influence of climate on tree growth was the same for both infected and healthy trees.     

Comparative proteomic analysis of kernel proteins of two high amylose transgenic durum wheat lines obtained by biolistic and Agrobacterium-mediated transformations

In this work, we compared the proteome of mature and immature kernels of transgenic and untransformed durum wheat lines in which genes of the starch branching enzymes of class II (SBEIIa) were silenced by RNA interference using two different methods of genetic transformation. The comparative analysis of granule bound and metabolic protein fractions of Svevo and Ofanto and their derived transgenic lines highlighted twenty four and thirty three differentially accumulated spots, respectively, in the line MJ16-112 (obtained by biolistic transformation of Svevo) and in the line A431_4p1a (obtained by Agrobacterium-mediated transformation of Ofanto).     

West Nile Virus in North‐Eastern Italy, 2011: Entomological and Equine IgM‐Based Surveillance to Detect Active Virus Circulation

Since 2008, West Nile Virus (WNV) has expanded its range in several Italian regions, and its yearly recurrence suggests the virus may have become endemic in some areas. In 2011, a new plan based also on the detection of IgM antibodies was implemented in the north‐eastern Italian regions of Veneto and Friuli Venezia Giulia, aiming to early detect WNV infections in areas where the virus had already circulated during the previous summers, and in adjacent zones. From July to November 2011, 1880 sera from 521 equine premises were screened by a commercial IgM capture ELISA. Mosquitoes were captured by CDC‐CO₂ traps at 61 locations in the two regions. Collected mosquitoes were identified, pooled by species/date/location and examined by real‐time RT‐PCR and sequencing. Passive surveillance was carried out on clinically affected horses and non‐migratory wild birds found dead. IgM sero‐positive equines were detected in 19 holdings, five in the area with WNV circulation (AWC) and 14 in the surveillance area (SA); 10 more horse premises tested positive to further serological controls within 4 km of the positive holdings. A total of 85 398 mosquitoes of 15 species were collected and 2732 pools examined. Five Culex pipiens pools tested positive for the presence of WNV. Passive surveillance on non‐migratory wild birds allowed detection of the virus only in one found dead collared dove (Streptopelia decaocto), of 82 birds sampled. The WNV belonged to the lineage 2, which had been isolated for the first time in Italy earlier in 2011. By the first week of October, nine human cases had been confirmed in the same area. The implementation of a protocol combining IgM screening of horses with surveillance on mosquito vectors proved to be valuable for early detecting WNV circulation.     

WHOLE BODY COMPUTED TOMOGRAPHIC CHARACTERISTICS OF SKELETAL AND CARDIAC MUSCULAR METASTATIC NEOPLASIA IN DOGS AND CATS

Muscular metastatic neoplasia has been reported to be rare in domestic animals, however previous studies were based primarily on necropsy findings. The purpose of this retrospective study was to describe whole body computed tomography (CT) characteristics of confirmed muscular metastases in a cohort of dogs and cats presented for oncology evaluation. Medical records of 1201 oncology patients were reviewed. Included animals underwent pre and postcontrast whole body CT, and CT‐guided tru‐cut biopsy or fine needle aspiration of one or more metastatic lesions. Twenty‐one dogs and six cats met inclusion criteria, representing 2.08% of all canine oncology patients and 3.1% of all feline oncology patients. Mean age was 9.6 years. Postcontrast CT characteristics included well‐demarcated, oval‐to‐round lesions with varying enhancement patterns: ring enhancing (n = 16), heterogeneously enhancing (n = 8), or homogeneously enhancing (n = 5). Five animals showed concurrent and varying nodular patterns. In seven cases (five dogs and two cats), one single muscular nodule was observed. In 20 cases, two or more lesions were observed. In two cases, cardiac hypodense nodules were observed in the postcontrast CT, while appearing isodense in the precontrast study. Necropsy confirmed neoplasia in both of them. Locations of muscular metastases included epaxial/paraspinal muscles of the cervical, thoracic, and lumbar spine (n = 18), superficial muscles of the thoracic wall (n = 13), scapular/shoulder region (n = 3), hind limb (n = 3), and abdominal wall muscles (n = 1). Findings supported the use of pre and postcontrast whole body CT for oncologic staging in dogs and cats, especially for primary tumors characterized by a high metastatic rate.     

Ultrasound-guided retrieval of plant awns

In dogs, surgical removal of plant awns causing a foreign body granuloma or abscess may be challenging. The inability to localize the foreign body during surgical removal often leads to abscess recurrence. In this report, we describe ultrasound-guided retrieval as an alternative to standard surgical retrieval in animals where the plant awn can be identified sonographically. This procedure was used in six dogs with a superficial abscess due to a plant awn, and in all dogs the foreign body was successfully retrieved with Hartmann forceps. No complication was observed and no abscess recurred. Minimally invasive ultrasound-guided plant awn retrieval appears to be a safe effective method of retrieving superficially located plant awns in the dog.     

Relative skeletal distribution of proliferating marrow in the adult dog determined using 3′‐deoxy‐3′‐[18F]fluorothymidine

3′‐deoxy‐3′‐[¹⁸F]fluorothymidine (¹⁸FLT) is a radiopharmaceutical tracer used with positron emission tomography (PET), often in combination with computed tomography (CT), to image DNA synthesis, and thus, cellular proliferation. Characteristic accumulation of the tracer within haematopoietic bone marrow provides a noninvasive means to assess marrow activity and distribution throughout the living animal. The present study utilizes three‐dimensional analysis of ¹⁸FLT‐PET/CT scans to quantify the relative skeletal distribution of active marrow by anatomic site in the dog. Scans were performed on six healthy, adult (3–6 years of age), mixed‐breed dogs using a commercially available PET/CT scanner consisting of a 64‐slice helical CT scanner combined with an integrated four ring, high‐resolution LSO PET scanner. Regions of interest encompassing 11 separate skeletal regions (skull, cervical vertebral column, thoracic vertebral column, lumbar vertebral column, sacrum, ribs, sternum, scapulae, proximal humeri, ossa coxarum, and proximal femora) were manually drawn based on CT images and thresholded by standardized uptake value to delineate bone marrow activity. Activity within each skeletal region was then divided by the total skeletal activity to derive the per cent of overall marrow activity within an individual site. The majority of proliferative marrow was located within the vertebral column. Of the sites traditionally accessed clinically for marrow sampling, the proximal humerus contained the largest percentage, followed by the ossa coxarum, proximal femur, and sternum, respectively. This information may be used to guide selection of traditional marrow sampling sites as well as inform efforts to spare important sites of haematopoiesis in radiation therapy planning.     

EFFECT OF SEDATION ON CONTRAST‐ENHANCED ULTRASONOGRAPHY OF THE SPLEEN IN HEALTHY DOGS

Contrast‐enhanced ultrasound of the spleen enables the dynamic assessment of the perfusion of this organ, however, both subjective and quantitative evaluation can be strongly influenced by sedative agent administration. The purpose of this prospective, experimental study was to test effects of two sedative agents on splenic perfusion during contrast‐enhanced ultrasound of the spleen in a sample of healthy dogs. Contrast‐enhanced ultrasound of the spleen was repeated in six healthy Beagles following a cross‐over study design comparing three protocols: awake, butorphanol 0.2 mg/Kg intramuscular (IM), and dexmedetomidine 500 μg/m² IM. After intravenous injection of a phospholipid stabilized sulfur hexafluoride microbubble solution (SonoVue®, Bracco Imaging, Milano, Italy), the enhancement intensity and perfusion pattern of the splenic parenchyma were assessed and perfusion parameters were calculated. Normal spleen was slightly heterogeneous in the early phase, but the parenchyma was homogeneous at a later phase. Sedation with butorphanol did not modify perfusion of the spleen. Dexmedetomidine significantly reduced splenic enhancement, providing diffuse parenchymal hypoechogenicity during the entire examination. Measured parameters were significantly modified, with increased arrival time (AT; (< 0.0001) and time to peak (TTP; P < 0.0001), and decreased peak intensity (PI; P = 0.0108), wash‐in (P = 0.0014), and area under the curve (AUC; P = 0.0421). Findings supported the use of butorphanol and contraindicated the use of dexmedetomidine as sedatives for splenic contrast ultrasound procedures in dogs. Short‐term and diffuse heterogeneity of the spleen in the early venous phase was determined to be a normal finding.