The prevalence of Salmonella infection was determined in a group of spur-thighed tortoises (Testudo graeca) seized during two smuggling attempts and in a population of captive Hermann's tortoises (Testudo hermanni) sheltered in a wildlife rescue centre. Salmonella spp. was isolated in 81 of 220 (36.8%) and in 17 of 67 (25.4%) cloacal swabs collected from the T. graeca and T. hermanni tortoises respectively. Overall, a total of 21 different Salmonella serotypes were found. Some of these serotypes are common to terrestrial chelonians while others have never been reported. All cultured serotypes were non-typhoidal but nonetheless many of these have been previously reported as source of human outbreaks of reptile-related salmonellosis. Eighty-two per cent and 5.3% of the isolates were resistant to two and three anti-microbial agents respectively. However, the isolates were highly susceptible to the anti-microbials of choice for the treatment of salmonellosis such as cephalosporins and fluoroquinolones. Our findings confirm that tortoises can be considered a reservoir for Salmonella and that care should be employed when handling and breeding these animals. Tight surveillance should be enforced to avoid illegal importation and prevent the trading of live tortoises, carriers of zoonotic pathogens.

The issue of whether market fish can be involved in the transmission of Toxoplasma gondii in the marine environment is highly debated since toxoplasmosis has been diagnosed frequently in cetaceans stranded along the Mediterranean coastlines in recent times. To support the hypothesis that fishes can harbour and effectively transmit the parasite to top‐of‐the‐food‐chain marine organisms and to human consumers of fishery products, a total of 1,293 fishes from 17 species obtained from wholesale and local fish markets were examined for T. gondii DNA. Real‐time PCR was performed in samples obtained by separately pooling intestines, gills and skin/muscles collected from each fish species. Thirty‐two out of 147 pooled samples from 12 different fish species were found contaminated with T. gondii DNA that was detected in 16 samples of skin/muscle and in 11 samples of both intestine and gills. Quantitative analysis of amplified DNA performed by both real‐time PCR and digital PCR (dPCR) confirmed that positive fish samples were contaminated with Toxoplasma genomic DNA to an extent of 6.10 × 10−2 to 2.77 × 104 copies/ml (quantitative PCR) and of 1 to 5.7 × 104 copies/ml (dPCR). Fishes are not considered competent biological hosts for T. gondii; nonetheless, they can be contaminated with T. gondii oocysts flowing via freshwater run‐offs (untreated sewage discharges, soil flooding) into the marine environment, thus acting as mechanical carriers. Although the detection of viable and infective T. gondii oocysts was not the objective of this investigation, the results here reported suggest that fish species sold for human consumption can be accidentally involved in the transmission route of the parasite in the marine environment and that the risk of foodborne transmission of toxoplasmosis to fish consumers should be further investigated.

BackgroundThe serotonergic system is associated with numerous brain functions, including the resetting of the mammalian circadian clock. The synthesis and metabolism of 5-HT in the brain increases in response to exercise and is correlated with high levels of blood-borne tryptophan (TRP). The present investigation was aimed at testing the existence of a daily rhythm of TRP and 5-HT in the blood of athletic horses.MethodsBlood samples from 5 Thoroughbred mares were collected at 4-hour intervals for 48 hours (starting at 08:00 hours on day 1 and finishing at 4:00 on day 2) via an intravenous cannula inserted into the jugular vein. Tryptophan and serotonin concentrations were assessed by HPLC. Data analysis was conducted by one-way repeated measures analysis of variance (ANOVA) and by the single cosinor method.ResultsANOVA showed a highly significant influence of time both on tryptophan and on serotonin, in all horses, on either day, with p values < 0.0001. Cosinor analysis identified the periodic parameters and their acrophases (expressed in hours) during the 2 days of monitoring. Both parameters studied showed evening acrophases.ConclusionThe results showed that serotonin and tryptophan blood levels undergo nycthemeral variation with typical evening acrophases. These results enhance the understanding of the athlete horse's chronoperformance and facilitate the establishment of training programs that take into account the nycthemeral pattern of aminoacids deeply involved in the onset of central fatigue.

We report a rapid and reliable method for the detection of Toxoplasma gondii in meat and animal tissues based on real-time polymerase chain reaction (PCR). Samples were collected from cattle, small ruminants, horses, and pigs raised or imported into Sicily, Italy. All DNA preparations were assayed by real-time PCR tests targeted to a 98-bp long fragment in the AF 529-bp repeat element and to the B1 gene using specific primers. Diagnostic sensitivity (100%), diagnostic specificity (100%), limit of detection (0.01 pg), efficiency (92-109%), and precision (mean coefficient of variation = 0.60%), repeatability (100%), reproducibility (100%), and robustness were evaluated using 240 DNA extracted samples (120 positives and 120 negative as per the OIE nested PCR method) from different matrices. Positive results were confirmed by the repetition of both real-time and nested PCR assays. Our study demonstrates the viability of a reliable, rapid, and specific real-time PCR on a large scale to monitor contamination with Toxoplasma cysts in meat and animal specimens. This validated method can be used for postmortem detection in domestic and wild animals and for food safety purposes.

The authors investigated mammary blood flow by means of a pulsed wave-Doppler ultrasonographic technique in ewes during different production phases. Three groups (Ga, Gb, and Gc, five animals in each) of Comisana sheep were used in the experiment. Ga sheep were milked twice a day by means of a milking machine; Gb sheep nursed their lambs, and group Gc was made up of dry sheep. Recordings of systolic and diastolic pressure of the left and right lateral mammary veins (SSLMV and DSLMV) and arteries (SSLMA and DSLMA) were carried out by means of a Pulse Doppler system. Recordings for Ga were taken before and after milking in the morning and in the afternoon. In Gb and Gc sheep, recordings were taken only in the morning and in the afternoon. Statistical analysis was carried out of the mean values of the recordings taken as mentioned above and the Student's t-test for paired data was used. A statistically significant difference was found in Ga recordings carried out before and after morning (0.05 < P < 0.001) and afternoon (0.05 < P < 0.01) milking. Ga morning mean flow speeds were between 7.