Copromicroscopic and molecular assays for the detection of cancer-causing parasitic nematode Spirocerca lupi

Spirocerca lupi (Nematoda, Spirurida) is a life-threatening parasitic nematode of dogs that is presently emerging in several countries. Nonetheless, canine spirocercosis is neglected and underestimated, mainly due to diagnostic limitations inherent to clinico-pathologic, diagnostic imaging and laboratory methodologies. Given the significant benefit of improved diagnosis, the present work evaluated the reliability of a recently described copromicroscopic approach, the FLOTAC technique, as well as a PCR-based assay with that of traditional coproscopic techniques to diagnose S. lupi infection. Ninety-four faecal field samples were collected from two endemic areas (i.e. 29 and 65 from Kenya and Israel, respectively) and processed using different coproscopic examination techniques. In particular, set I (Kenyan samples) comprised the modified flotation with Sheather's sugar solution and merthiolate-iodine-formalin technique, while set II (Israeli samples) comprised a flotation technique with zinc sulphate solution, a modified sugar flotation procedure and the FLOTAC method. All samples were also subjected to a semi-nested PCR protocol specific for a region internal to the mitochondrial cytochrome c oxidase subunit 1 gene of S. lupi. The coproscopic examinations showed low sensitivity and high variability, demonstrating the unreliability of the conventional methods for detecting S. lupi eggs. Nonetheless, the FLOTAC technique scored the highest number of positives and significantly higher number of S. lupi eggs per microscopic field compared to the other coproscopic methods. Additionally, of the coproscopically negative samples, 9 (45%) Kenyan and 21 (38.2%) Israeli samples scored molecularly positive using the PCR-based approach. The potential implications and perspectives for canine spirocercosis of these coproscopic and molecular diagnostic methodologies evaluated herein are discussed.     

Effects of compost and defatted oilseed meals as sustainable organic fertilisers on cardoon (Cynara cardunculus L.) production in the Mediterranean basin

Cardoon (Cynara cardunculus L.) is considered as one of the most suitable energy crops for Southern Europe. The aim of this work was to outline the effects of organic fertilisers on the productivity and the global warming potential (GWP) on cardoon production. Six fertilisers (N 100 kg ha^?1, N 50 kg ha^?1, Compost 30 t ha^?1, Compost 15 t ha^?1 + N 25 kg ha^?1, 3 t ha^?1 of defatted oilseed meals of sunflower, 3 t ha^?1 of defatted oilseed meals of Brassica carinata), and unfertilised control, were evaluated on two cultivars (‘Gobbo di Nizza’ and ‘Altilis 41’) in a split-plot experiment. Defatted oilseed meal of sunflower recorded higher total dry weight (+10%), seed yield (+17%), nitrogen use efficiency (+14%) and better GWP (?66%) compared to the other organic fertilisers and performing as well as N 100 kg ha^?1. Altilis 41 cultivar showed the highest aboveground total dry weight (10 t ha^?1 y^?1), seed yield (1.7 t ha^?1 y^?1), stalk dry weight (7 t ha^?1 y^?1) and head dry weight (3 t ha^?1 y^?1). Our results highlighted that by combining suitable cultivar and fertilisation strategies, it could be possible to increase the production sustainability of C. cardunculus.     

Unicentric extranodal lymphoma of the upper eyelid conjunctiva in a dog

A case of conjunctival lymphoma in a 4-year-old Siberian husky is reported. A large red mass protruding from the conjunctiva of the upper eyelid of the right eye was present. Irritation, blepharospasm and epiphora were revealed on initial ophthalmic examination. After anti-inflammatory treatment, surgery was performed. Histologically, the mass consisted of large polygonal cells with a high nucleus to cytoplasm ratio, moderate amounts of slightly eosinophilic cytoplasm, pleomorphic nuclei with vesicular chromatin and prominent multiple nucleoli. Mitotic figures were frequent. Approximately 70% of the neoplastic cells were CD3 positive and CD79alpha negative. On the basis of histologic and immunohistochemical findings, a diagnosis of intermediate-grade, diffuse large-cell lymphoma with T-cell immunophenotype was made. The surgical area healed uneventfully and, although chemotherapy was not received, after 12 months the dog exhibited no recurrence. To the authors' knowledge, this is the first report of unicentric extranodal conjunctival lymphoma in a dog.     

Seroprevalence of Neospora spp. in asymptomatic horses in Italy

One hundred and fifty horses from Italy were tested for serum antibodies to Neospora caninum by indirect fluorescence antibody test (IFAT). Antibodies were present in 42 horses (28%). The positive titres were: 1:50 (16/150-10.5%), 1:100 (26/150-17%), 1:200 (8/150-5.3%) and 1:800 (1/150-0.6%). No neurological anomalies were observed in the seropositive horses. This is the first serologic survey for Neospora spp. antibodies performed on horses born and raised in Italy.     

Report in Europe of nasal myiasis by Rhinoestrus spp. in horses and donkeys: seasonal patterns and taxonomical considerations

Two species of Rhinoestrus (i.e. Rhinoestrus purpureus (Brauer) and Rhinoestrus usbekistanicus Gan) cause nasal myiasis in horses, donkeys and zebras. In the past 15 years myiasis caused by R. purpureus has been reported in Egypt and by R. usbekistanicus in Senegal and Niger, both in horses and in donkeys. With the aim to investigate the presence of this myiasis in autochthonous horses and donkeys from southern Italy and to study the seasonal trend of larval infection, 212 native horses were necropsied in two slaughterhouses in the Apulia region (site A) from January to November 2003, and 120 native horses and two donkeys in one slaughterhouse in Sicily (site B) from January to October 2003. Thirteen of 212 and 5 of 120 horses examined from sites A and B, respectively, were infected by nasal bot fly larvae. Both donkeys examined were positive. Two hundred and thirteen larvae, representing all stages, were collected from the throat region, the turbinates and beyond the lamina cribrosa of the ethmoid bone, in the cerebral cavity. Third larval stages were retrieved from April to September with the highest mean burden in site A in May and in July in site B. The simultaneous presence all three larval stages at site B suggests the existence of two or more overlapping generations. Larvae were identified on the basis of peritreme structures and arrangement of the spines on the dorsal surface of the third segment. Some L3 collected from sites A and B presented morphological characteristics of both R. usbekistanicus and R. purpureus. The presence of mixed features on the L3 of Rhinoestrus collected from the same animals may be explained by the existence of a single species of Rhinoestrus presenting different morphotypes or of two species of Rhinoestrus which are both present in southern Italy since this area is midway between eastern and African countries where these two species have been reported.     

Intraoperative Analgesic Effect of Intrafunicular Lidocaine Injection During Orchiectomy in Isoflurane-Anesthetized Martina Franca Donkeys

This study aimed to evaluate the analgesic effect of intrafunicular lidocaine during orchiectomy in isoflurane-anesthetized donkeys. For this purpose, 10 adult healthy donkeys were chosen from Martina Franca donkey population. Each donkey underwent two surgical procedures of monolateral orchiectomy under general anesthesia. Starting isoflurane vaporizer setting was 1.5% to maintain a light plane of anesthesia. Regional anesthesia was performed by injecting 10 mL of 2% lidocaine plus adrenalin or an equivalent volume of saline solution into the spermatic cord. According to what was injected into the funiculus before the surgical procedure, each donkey was once assigned to the group L (lidocaine) and once to the group S (saline). End-tidal isoflurane and standard physiological parameters were measured. Compared with groups, monolateral orchiectomy increased mean heart rate in group S; during surgical procedure, the end-tidal isoflurane was significantly lower in group L. No differences were found regarding mean arterial blood pressure, respiratory rate, recovery quality, and metabolic parameters between groups during procedures. In isoflurane-anesthetized donkeys, intrafunicular injection of lidocaine before castration appears to decrease intraoperative nociception and significantly reduces the concentration of the volatile agent to obtain a sufficient surgical anesthesia.     

IMAGING DIAGNOSIS—TRANSESOPHAGEAL ULTRASOUND‐GUIDED REMOVAL OF A MIGRATING GRASS AWN FOREIGN BODY IN A DOG

A 3‐year‐old English Setter dog was presented for an acute onset of coughing. Tracheobronchoscopic examination allowed localization and removal of one grass awn foreign body. A second migrated grass awn was suspected to be present in the left caudal lung lobe. Transesophageal ultrasound revealed an area of pulmonary consolidation in the dorsomedial portion of left caudal lobe and a linear hyperechoic structure consistent with a grass awn foreign body within the area of consolidation. Transesophageal ultrasonography was also used to provide anatomical landmarks that facilitated successful thoracoscopic removal of the foreign body.     

Hereditary early-onset Parkinson's disease caused by mutations in PINK1

Parkinson's disease (PD) is a neurodegenerative disorder characterized by degeneration of dopaminergic neurons in the substantia nigra. We previously mapped a locus for a rare familial form of PD to chromosome 1p36 (PARK6). Here we show that mutations in PINK1 (PTEN-induced kinase 1) are associated with PARK6. We have identified two homozygous mutations affecting the PINK1 kinase domain in three consanguineous PARK6 families: a truncating nonsense mutation and a missense mutation at a highly conserved amino acid. Cell culture studies suggest that PINK1 is mitochondrially located and may exert a protective effect on the cell that is abrogated by the mutations, resulting in increased susceptibility to cellular stress. These data provide a direct molecular link between mitochondria and the pathogenesis of PD.     

Seismic evidence for water deep in Earth's upper mantle

Water in the deep upper mantle can influence the properties of seismic discontinuities in the mantle transition zone. Observations of converted seismic waves provide evidence of a 20- to 35-kilometer-thick discontinuity near a depth of 410 kilometers, most likely explained by as much as 700 parts per million of water by weight.     

Identification of a universal Group B streptococcus vaccine by multiple genome screen

Group B Streptococcus (GBS) is a multiserotype bacterial pathogen representing a major cause of life-threatening infections in newborns. To develop a broadly protective vaccine, we analyzed the genome sequences of eight GBS isolates and cloned and tested 312 surface proteins as vaccines. Four proteins elicited protection in mice, and their combination proved highly protective against a large panel of strains, including all circulating serotypes. Protection also correlated with antigen accessibility on the bacterial surface and with the induction of opsonophagocytic antibodies. Multigenome analysis and screening described here represent a powerful strategy for identifying potential vaccine candidates against highly variable pathogens.