FIRST RESULTS ON THE PRESENCE AND THE MOLECULAR CHARACTERIZATION OF ANISAKID NEMATODES IN MARINE FISH CAUGHT OFF NORTHERN SARDINIA

Anisakidosis is a parasitic zoonosis caused by nematodes of the family Anisakidae, belonging to the genera Anisakis, Contracaecum and Pseudoterranova. Molecular studies have shown that Anisakis larvae comprise a number of sibling species, which have different genetic structures, hosts and geographical distribution. A great variety of fish species can harbour infectious third stage larvae of this nematode. The preliminary results of a study carried out to evaluate the occurrence of this parasite in commercial fish caught off northern Sardinia are herein reported. From October 2008 to November 2009, 599 specimens of 8 commercial fish species were examined for anisakid larvae through visual inspection of body cavity and peptic digestion of the muscle. Isolated Anisakis sp. larvae were observed at light microscope and identified as Type I or Type II (sensu Berland, 1961). Out of 599 fish examined, 239 (40%) were infected by 1187 anisakid larvae, belonging to the genera Anisakis (1169 Type I and 18 Type II) and Hysterothylacium (692). The molecular identification of Anisakis spp. was carried out on a subsample of 30% of Type I larvae and all Type II larvae. Specimens were firstly examined using a species-specific PCR, with primers designed for Anisakis pegreffii (APEF) and Anisakis physeteris (APHF), and ITS-2 of nuclear rDNA. The results were confirmed by the analysis of the ITS region of nuclear rDNA (ITS-1, 5.8S and ITS-2) using the restriction enzymes HinfI and HhaI in PCR-RFLP. Type I larvae examined were all identified as A. pegreffii, and Type II were all A. physeteris. This is the first contribution to the epidemiology and molecular characterization of Anisakis spp. in commercial fish caught off Sardinia

Milk cathelicidin and somatic cell counts in dairy goats along the course of lactation

This research communication reports the evaluation of cathelicidin in dairy goat milk for its relationship with the somatic cell count (SCC) and microbial culture results. Considering the limited performances of SCC for mastitis monitoring in goats, there is interest in evaluating alternative diagnostic tools. Cathelicidin is an antimicrobial protein involved in innate immunity of the mammary gland. In this work, half-udder milk was sampled bimonthly from a herd of 37 Alpine goats along an entire lactation and tested with the cathelicidin ELISA together with SCC and bacterial culture. Cathelicidin and SCC showed a strong correlation (r = 0.72; n = 360 milk samples). This was highest in mid-lactation (r = 0.83) and lowest in late lactation (r = 0.61), and was higher in primiparous (0.80, n = 130) than in multiparous goats (0.71, n = 230). Both markers increased with stage of lactation, but cathelicidin increased significantly less than SCC. Inaddition, peak level in late lactation was lower for cathelicidin (5.05-fold increase) than for SCC (7.64-fold increase). Twenty-one (5.8%) samples were positive to bacteriological culture, 20 for coagulase-negative staphylococci and one for Streptococcus spp.; 18 of them were positive to the cathelicidin ELISA (85.71% sensitivity). Sensitivity of SCC >500 000 and of SCC >1 000 000 cells/ml was lower (71.43 and 23.81%, respectively). Therefore, the high correlation of cathelicidin with SCC during the entire lactation, along with its lower increase in late lactation and good sensitivity indetecting intramammary infection (IMI), indicate a potential for monitoring subclinical mastitis in dairy goats. However, based on this preliminary assessment, specificity should be improved (40.41% for cathelicidin vs. 54.57 and 67.85% for SCC >500 000 and >1 000 000 cells/ml, respectively). Therefore, the application of cathelicidin for detecting goat IMI will require further investigation and optimization, especially concerning the definition of diagnostic thresholds

Preliminary efficacy and safety data of nivolumab in never smoker patients with advanced squamous NSCLC: Experience from Italian sites participating in the Expanded Access Programme (EAP)

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A comparison of 4 different machine learning algorithms to predict lactoferrin content in bovine milk from mid-infrared spectra

peer-reviewedLactoferrin (LF) is a glycoprotein naturally present in milk. Its content varies throughout lactation, but also with mastitis; therefore it is a potential additional indicator of udder health beyond somatic cell count. Condequently, there is an interest in quantifying this biomolecule routinely. First prediction equations proposed in the literature to predict the content in milk using milk mid-infrared spectrometry were built using partial least square regression (PLSR) due to the limited size of the data set. Thanks to a large data set, the current study aimed to test 4 different machine learning algorithms using a large data set comprising 6,619 records collected across different herds, breeds, and countries. The first algorithm was a PLSR, as used in past investigations. The second and third algorithms used partial least square (PLS) factors combined with a linear and polynomial support vector regression (PLS + SVR). The fourth algorithm also used PLS factors, but included in an artificial neural network with 1 hidden layer (PLS + ANN). The training and validation sets comprised 5,541 and 836 records, respectively. Even if the calibration prediction performances were the best for PLS + polynomial SVR, their validation prediction performances were the worst. The 3 other algorithms had similar validation performances. Indeed, the validation root mean squared error (RMSE) ranged between 162.17 and 166.75 mg/L of milk. However, the lower standard deviation of cross-validation RMSE and the better normality of the residual distribution observed for PLS + ANN suggest that this modeling was more suitable to predict the LF content in milk from milk mid-infrared spectra (R2v = 0.60 and validation RMSE = 162.17 mg/L of milk). This PLS +ANN model was then applied to almost 6 million spectral records. The predicted LF showed the expected relationships with milk yield, somatic cell score, somatic cell count, and stage of lactation. The model tended to underestimate high LF values (higher than 600 mg/L of milk). However, if the prediction threshold was set to 500 mg/L, 82% of samples from the validation having a content of LF higher than 600 mg/L were detected. Future research should aim to increase the number of those extremely high LF records in the calibration set

Diaphragmatic pacing stimulation in spinal cord injury: anesthetic and perioperative management

OBJECTIVE: The standard therapy for patients with high-level spinal cord injury is long-term mechanical ventilation through a tracheostomy. However, in some cases, this approach results in death or disability. The aim of this study is to highlight the anesthetics and perioperative aspects of patients undergoing insertion of a diaphragmatic pacemaker. METHODS: Five patients with quadriplegia following high cervical traumatic spinal cord injury and ventilator-dependent chronic respiratory failure were implanted with a laparoscopic diaphragmatic pacemaker after preoperative assessments of their phrenic nerve function and diaphragm contractility through transcutaneous nerve stimulation. ClinicalTrials.gov: NCT01385384. RESULTS: The diaphragmatic pacemaker placement was successful in all of the patients. Two patients presented with capnothorax during the perioperative period, which resolved without consequences. After six months, three patients achieved continuous use of the diaphragm pacing system, and one patient could be removed from mechanical ventilation for more than 4 hours per day. CONCLUSIONS: The implantation of a diaphragmatic phrenic system is a new and safe technique with potential to improve the quality of life of patients who are dependent on mechanical ventilation because of spinal cord injuries. Appropriate indication and adequate perioperative care are fundamental to achieving better results

MOLECULAR CHARACTERIZATION OF ANISAKID NEMATODES IN FISHES OF NORTHERN SARDINIAN SEA

The authors report results of analysis carried out during 2008-2010 for identification and molecular characterization of larval Anisakis nematodes isolated from fishes of the northern Sardinian sea