Agronomic and phytotoxicity test with biosolids from anaerobic CO-DIGESTION with temperature and micro-organism phase separation, based on sewage sludge, vinasse and poultry manure.

This study deals with energy and agronomic valorisation by anaerobic co-digestion with temperature and microorganism phase separation of sewage sludge, vinasse and poultry manure, with the aim of achieving an integral waste management, obtaining bioenergy and biofertilizer that returns nutrients to the soil in a natural way. The yields obtained were 40 mL H2/gVS and 391 mLCH4/gVS. The resulting effluent showed more than 98 % removal of E. coli and Total Coliforms, as well as total removal of Salmonella. The results obtained in the phytotoxicity tests showed that all the proportions studied had phytostimulant and phytonutrient properties, with 20 % having the highest germination index (GI) with mean values of 145.30 %. Finally, the agronomic trial carried out with strawberry crops (Fragaria sp.) showed that the addition of this biosolid has fertilising properties and can be used as an agronomic amendment, with an increase of 145 % in fresh weight and 102.5 % in dry weight, and fruit production doubled with respect to the control. The ANOVA statistical study corroborated that there were significant differences in crop growth when applying different proportions of biofertilizer in the fertilizer. Therefore, these results show that this technology is promising and would contribute environmentally, socially and economically to the transfer towards a circular economy model.

Pathological changes and viral antigen distribution in tissues of Iberian hare (Lepus granatensis) naturally infected with the emerging recombinant myxoma virus (ha-MYXV).

BACKGROUND: A cross-species jump was confirmed in 2018, when a novel recombinant myxoma virus (MYXV) (ha-MYXV) caused high mortality in Iberian hare (Lepus granatensis) in the Iberian Peninsula. METHOD: The aim of this study was to evaluate the main lesions, tissular distribution and target cells of ha-MYXV in Iberian hare. Gross postmortem examinations and histological and immunohistochemical studies to detect ha-MYXV were carried out in 28 animals that were confirmed as ha-MYXV positive by PCR. RESULTS: The main macroscopic lesions were bilateral blepharoconjunctivitis, epistaxis, intense congestion and oedema in several organs and some internal haemorrhages. Visible myxomas were not found. Histopathological examination revealed hyperplastic epidermis with predominant hyperkeratosis and myxoid matrix in the dermis. ha-MYXV-positive keratinocytes showed hydropic degeneration and cytoplasmic inclusion bodies. Alveolar oedema, interstitial pneumonia, dramatic lymphoid depletion in the spleen and necrosis in the liver and testis were observed. ha-MYXV was mainly detected in epithelial and myxoma cells in the skin, and also in macrophages, lymphocytes, fibroblasts and endothelial cells in several organs, as well as in hepatocytes and Leydig cells. LIMITATIONS: A non-homogeneous number of samples were included in all the animals. Future experimental studies with controlled variables are necessary. CONCLUSION: These findings correspond to an unusual form of myxomatosis, characterised by an acute or hyperacute presentation.

Spatiotemporal monitoring of myxomatosis in European wild rabbit (Oryctolagus cuniculus) in Spanish Mediterranean ecosystems.

A long-term active epidemiological surveillance programme was conducted to determine seroprevalence to myxoma virus (MYXV), infection prevalence and spatiotemporal patterns and factors associated with MYXV circulation in wild rabbits (Oryctolagus cuniculus) in Spanish Mediterranean ecosystems. A total of 2376 animals were sampled over four study periods: 2009-2012 (P1), 2012-2015 (P2), 2015-2018 (P3) and 2018-2021 (P4). Antibodies against MYXV were detected by a commercial indirect ELISA in 59.9% (1424/2376; 95% CI: 58.0-61.9) of wild rabbits. At least one seropositive animal was detected on 131 (96.3%) of 136 game estates sampled. MYXV infection was confirmed by PCR in 94 of 1063 (8.8%; 95% CI: 7.3-10.7) wild rabbits. Circulation of the novel recombinant MYXV (ha-MYXV) was not found in wild rabbits analysed during P4. Five statistically significant spatiotemporal clusters of high MYXV seroprevalence were identified using a Bernoulli model: one in P2 and four in P3. A generalized linear mixed model (GLMM) analysis identified sampling season (autumn), age (adult and juvenile), outbreaks of myxomatosis in the month prior to sampling, mean annual temperature, humidity and seropositivity to rabbit haemorrhagic disease virus as factors potentially linked with MYXV seropositivity. GLMM analysis identified outbreaks of myxomatosis in the month prior to sampling, MYXV seropositivity and presence of lesions compatible with myxomatosis as factors associated with MYXV infection. The results indicate high exposure, widespread but non-homogeneous distribution, and endemic circulation of MYXV in wild rabbit populations in southern Spain during the last decade. Prevalence of antibodies against MYXV showed fluctuations both within the year and over the study periods, revealing variations in the immunity of wild rabbit populations in Mediterranean ecosystems that could increase the risk of MYXV re-emergence in immunologically naïve populations. The present study highlights the importance of long-term surveillance to better understand the epidemiology of MYXV in wild lagomorphs.

Spatiotemporal monitoring of Cysticercus pisiformis in European wild rabbit (Oryctolagus cuniculus) in Mediterranean ecosystems in southern Spain.

Cysticercosis in wild lagomorphs is caused by Cysticercus pisiformis, the larval stage of Taenia pisiformis. Although previous studies have reported the presence of T. pisiformis in different wild carnivore species, information about the prevalence of C. pisiformis in their intermediate hosts is still very scarce. An epidemiological surveillance program was carried out to determine the prevalence and spatiotemporal patterns of cysticercosis in wild rabbits (Oryctolagus cuniculus) from Spanish Mediterranean ecosystems. A total of 2,923 animals were sampled in 164 hunting estates from Andalusia (southern Spain) during four study periods: 2009-2012 (P1), 2012-2015 (P2), 2015-2018 (P3) and 2018-2020 (P4). The presence of cysticerci was assessed by macroscopical examination and a subset of the collected parasites were molecularly identified by conventional PCR targeting the ITS-1 and 12S rRNA partial genes of Taenia spp. Risk factors associated with cysticercus infection were assessed by generalized estimating equation (GEE) analysis. A spatial statistical analysis was carried out using a Bernoulli model to identify statistically significant spatial clusters. Cysticercus infection was confirmed in 81 (2.8 %; 95 % CI: 2.2-3.4) rabbits. Cysticerci from 18 infected animals were molecularly identified as T. pisiformis. The GEE model showed the study period as the only risk factor associated with C. pisiformis infection in wild rabbits. Significantly higher prevalence was found in P2 (6.1 %; 95 % CI: 4.4-8.4) compared to the rest of the periods. At least one cysticerci-positive animal was detected in 41 (25.0 %; 95 % CI: 18.4-31.6) out of the 164 hunting estates. No statistically significant spatial clusters of high cysticercus prevalence were identified. Our results indicate an endemic circulation of C. pisiformis in wild rabbits in southern Spain. The spatial results highlight a widespread distribution of this parasite in their populations. Further studies should focus in determining which sympatric species may act as definitive hosts for T. pisiformis and the relevance of other potential intermediate host species (e.g. hares and rodents), as the relevance of wild rabbits in the sylvatic cycle of this cestode in Mediterranean ecosystems seems to be low.

Monitoring of emerging myxoma virus epidemics in Iberian hares (Lepus granatensis) in Spain, 2018-2020.

Myxomatosis is an infectious disease caused by the myxoma virus (MYXV), which has very high mortality rates in European wild rabbits (Oryctolagus cuniculus). While sporadic cases of myxomatosis have also been reported in some hare species, these lagomorphs are considered to have a low susceptibility to MYXV infection. In the present study, we describe the spatiotemporal evolution and main epidemiological findings of novel hare MYXV (ha-MYXV or MYXV-Tol) epidemics in Iberian hares (Lepus granatensis) in Spain. In the period 2018-2020, a total of 487 hares from 372 affected areas were confirmed to be MYXV-infected by PCR. ha-MYXV outbreaks were detected in most of the Spanish regions where the Iberian hare is present. The spatial distribution was not homogeneous, with most outbreaks concentrated in the southern and central parts of Spain. Consecutive outbreaks reported in the last two years suggest endemic circulation in Spain of this emerging virus. A retrospective study carried out just after the first epidemic period (2018-2019) revealed that the virus could have been circulating since June 2018. The number of outbreaks started to rise in July, peaked during the first half of August and October and then decreased sharply until January 2019. The apparent mean mortality rate was 55.4% (median: 70%). The results indicated high susceptibility of the Iberian hare to ha-MYXV infection, but apparent resistance in the sympatric hare species present in Spain and less infectivity in European rabbits. The novel ha-MYXV has had significant consequences on the health status of Iberian hare populations in Spain, which is of animal health and conservation concern. The present study contributes to a better understanding of ha-MYXV emergence and will provide valuable information for the development of control strategies. Further research is warranted to assess the impact of this emerging virus on wild lagomorph populations and to elucidate its ecological implications for Iberian Mediterranean ecosystems.

First outbreak of myxomatosis in Iberian hares (Lepus granatensis).

Myxomatosis is an infectious disease caused by myxoma virus (MYXV; genus Leporipoxvirus), which affects the European wild rabbit (Oryctolagus cuniculus) and sporadically brown hares (Lepus europaeus). Here, we describe the first outbreak of myxomatosis in Iberian hares (Lepus granatensis). Between mid-July and the end of September 2018, around 530 dead animals were detected in Iberian hare populations in southern Spain. The apparent mean mortality rate was 56.7%, and the estimated mean case fatality rate was 69.2%. Histopathological and molecular results confirmed MYXV infections in all hares analysed. To the authors' knowledge, this is the first myxomatosis outbreak causing a high mortality in hares and the first detailed characterization of a myxomatosis outbreak in the Iberian hare. The absence of cases in sympatric wild rabbits suggests differences in the susceptibility between both lagomorph species to the virus strain implicated in the outbreak. After the first case, the number of affected areas increased sharply affecting most of the Iberian Peninsula where the Iberian hare is present. Further studies are required to elucidate the origin of the implicated MYXV strain as well as to assess the impact of this outbreak on the Iberian hare populations.