Cyanobacterial dynamics and toxins concentrations in Lake Alto Flumendosa, Sardinia, Italy

Seasonal blooms of cyanobacteria (CB) are a typical feature of Lake Alto Flumendosa (Sardinia, Italy). The waters of this lake are used for drinking water supply, for agricultural and industrial uses, and fish farming activities. Since cyanotoxins are not monitored in edible organisms, diet could be a relevant route of human exposure. CB also represent a threat for the health of wild and domestic animals that use lake water for beverage. Therefore, to characterize the CB community and assess the risk for human and animal population, CB dynamic, mcyB+ fraction, and microcystins (MCs) concentration have been followed monthly for 18 months, in three stations. Results confirmed the presence of several toxigenic species. Planktothrix rubescens dominated between August 2011 and April 2012 (3.5×106 cells L-1), alternating with Woronichinia naegeliana (8×106 cells L-1) and Microcystis botrys (9×105 cells L-1). Dolichospermum planctonicum was always present at low densities (104 cells L-1). MCs were detected, at values well below the 1 µg L-1 threshold of WHO for drinking water. The molecular analysis of mcyB gene for P. rubescens indicated the presence of a persistent toxic population (average 0.45 mcyB/16S rDNA). Highly significant linear regressions were found between P. rubescens and the sum of the demethylated MC variants, and between M. botrys and the sum of MC-LR and MC-LA, also when co-occurring, suggesting that these two species were responsible for different MC patterns production. The regression lines indicated a quite stable MC cell quota. However, in some spotted samples very different values were obtained for both MC concentrations and cell quota (from 10-fold lower to 30-40-fold higher than the 'average') showing an unexpected significant variability in the rate of toxin production. The relatively low cell densities during the monitoring period is consistent with the low-to absent MC contamination level found in trout muscle; however, the analytical method was affected by low recovery, probably due to MC-protein binding. Our results show that, during the study period, no risk of exposure for the human and animal population occurred. However, the persistence of a complex CB community characterised by a significant toxic fraction suggests the need for periodic monitoring activity. Particularly, the hidden deep summer P. rubescens blooms, located where water is taken for drinking water supply, and M. botrys, able to produce the most toxic MC variants with high cell quota, should be kept under control. The documentation and interpretation of sudden changes in toxins concentrations deserve special attention. This is particularly relevant in proximity of fish farming plants and water catchment sites

Cyanobacterial dynamics and toxins concentrations in Lake Alto Flumendosa, Sardinia, Italy

Seasonal blooms of cyanobacteria (CB) are a typical feature of Lake Alto Flumendosa (Sardinia, Italy). The waters of this lake are used for drinking water supply, for agricultural and industrial uses, and fish farming activities. Since cyanotoxins are not monitored in edible organisms, diet could be a relevant route of human exposure. CB also represent a threat for the health of wild and domestic animals that use lake water for beverage. Therefore, to characterize the CB community and assess the risk for human and animal population, CB dynamic, mcyB+ fraction, and microcystins (MCs) concentration have been followed monthly for 18 months, in three stations. Results confirmed the presence of several toxigenic species. Planktothrix rubescens dominated between August 2011 and April 2012 (3.5×106 cells L-1), alternating with Woronichinia naegeliana (8×106 cells L-1) and Microcystis botrys (9×105 cells L-1). Dolichospermum planctonicum was always present at low densities (104 cells L-1). MCs were detected, at values well below the 1 µg L-1 threshold of WHO for drinking water. The molecular analysis of mcyB gene for P. rubescens indicated the presence of a persistent toxic population (average 0.45 mcyB/16S rDNA). Highly significant linear regressions were found between P. rubescens and the sum of the demethylated MC variants, and between M. botrys and the sum of MC-LR and MC-LA, also when co-occurring, suggesting that these two species were responsible for different MC patterns production. The regression lines indicated a quite stable MC cell quota. However, in some spotted samples very different values were obtained for both MC concentrations and cell quota (from 10-fold lower to 30-40-fold higher than the ‘average’) showing an unexpected significant variability in the rate of toxin production. The relatively low cell densities during the monitoring period is consistent with the low-to absent MC contamination level found in trout muscle; however, the analytical method was affected by low recovery, probably due to MC-protein binding. Our results show that, during the study period, no risk of exposure for the human and animal population occurred. However, the persistence of a complex CB community characterised by a significant toxic fraction suggests the need for periodic monitoring activity. Particularly, the hidden deep summer P. rubescens blooms, located where water is taken for drinking water supply, and M. botrys, able to produce the most toxic MC variants with high cell quota, should be kept under control. The documentation and interpretation of sudden changes in toxins concentrations deserve special attention. This is particularly relevant in proximity of fish farming plants and water catchment sites

Angiostrongylus chabaudi Biocca, 1957: A new parasite for domestic cats?

Background: Natural infection with a species of Angiostrongylus has been reported only once in wildcats from central Italy by Biocca in 1957. The causative species of this infection was identified as Angiostrongylus chabaudi. Following this report, this parasite had never been found in either wild or domestic cats. Findings: The lungs and the pulmonary arteries of an adult female cat (Felis silvestris catus), road-killed in Sardinia, Italy, were macroscopically examined and dissected under a light microscope for the presence of parasites. A slender nematode was detected and its morphometrical features were consistent with those of A. chabaudi. Morphological data were supplemented by sequencing of the partial cytochrome oxidase c subunit 1 (cox1) gene, as well as the internal transcribed spacer 2 (ITS2) of the rDNA. Nucleotide sequences displayed 99% homology with the ITS2 sequence [GenBank KM216825.1] of a specimen of Angiostrongylus sp. recovered recently from the pulmonary artery of a wildcat in Germany and 91% with cox1 sequence [GenBank GU138118.1] of Angiostrongylus vasorum. Conclusion: The results of the present study indicate, for the first time, that A. chabaudi may also infect domestic cats, and thus should be considered in the diagnosis of metastrongyloid species infecting their cardio-pulmonary system

The illegal rearing and slaughtering of pigs in the wild on the Mediterranean island of Sardinia favor an increase in the biomass of Trichinella britovi in wild boars (Sus scrofa) but do not affect the serological prevalence of infection

Abstract Background Worms of the nematode genus Trichinella are zoonotic pathogens with a worldwide distribution. The first report of Trichinella on the Mediterranean island of Sardinia was for Trichinella britovi, one of the four species of this genus circulating in Europe, which was identified in 2005 following an outbreak of trichinellosis in humans due to the consumption of pork from pigs reared in the wild. Since then, T. britovi larvae have been repeatedly isolated from free-ranging pigs, foxes (Vulpes vulpes) and wild boars (Sus scrofa) sampled in the central-eastern region of the island (Orgosolo municipality), but have never been isolated from samples from other areas of the island. The aim of this study was to investigate the parasitological and serological prevalence of T. britovi infection in wild boars in Sardinia over space [eight wild boar hunting management units (HMUs)] and time (seven wild boar hunting seasons). Methods Muscle and serum samples of boars killed in the 2014–2015 to 2020–2121 hunting seasons were collected from eight HMUs of central and south-western Sardinia. Trichinella sp. larvae were detected by artificial digestion of predilection muscles. A total of 4111 serum samples of wild boar were collected from the investigated HMUs and tested by enzyme-linked immunosorbent assay as a screening test and by western blot as a confirmatory test using excretory/secretory antigens. Results Trichinella britovi muscle larvae were detected in six (0.03%) of the 17,786 wild boars tested. All of the Trichinella sp.-positive wild boars had been hunted in Orgosolo municipality (central-eastern area of the island), except for one, hunted in a neighboring municipality. An overall serological prevalence of 3.8% (95% confidence interval, 3.3–4.5) was detected by western blot. No statistical differences were detected between the HMUs where T. britovi larvae were detected in wild boars, foxes, and free-ranging pigs and those where wild boars, foxes and free-ranging pigs tested negative. Conclusions The serological prevalence did not vary between the wild boar populations in which the larval load was detectable by artificial digestion (Orgosolo municipality) and those in which the larval load was below the detection limit. Furthermore, the serological prevalence of anti-Trichinella immunoglobulin G in the wild boar populations remained constant during the study period, which covered seven wild boar hunting seasons. As the transmission events (i.e., the serological prevalence) are stable, the high biomass of the parasite in Orgosolo municipality can only have arisen as a consequence of factors independent of its natural cycle, i.e., the presence of a high number of free-ranging pigs, and the concomitant presence of African swine fever, due to illegal pig slaughtering in the field. This epidemiological situation suggests that the natural cycle of T. britovi may be influenced by inappropriate pig husbandry and slaughtering practices. Graphical Abstrac