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Global fish and seafood consumption is increasing annually, frequently leading to the emergence of food fraud, mainly related to mislabeling and adulteration like, for example, the use of illicit/unauthorized food additives to mask or delay fish spoilage. Among the available diagnostic tools for control purposes, spectroscopic techniques have often been proposed to identify these kinds of illicit practices in fish and seafood products. The presented study aims to test two cheap and portable near infrared (NIR) spectrometers, a handheld MicroNIR and a pocket-sized SCiO, to uncover use of the illicit food additive Cafodos, a mixture of sodium citrate and hydrogen peroxide used to preserve some fish characteristics (like smell, color, na dtexture). The NIR spectroscopy in combination with chemometric approaches, allowed the successfully classification of (81–100%) samples of sea bass (Dicentrarchus labrax) treated with Cafodos. The study highlights the potential of this technique that, by not requiring pre-treatment of samples with further reagents, is cheaper and safer for the environment. In conclusion, the study confirmed the potential of portable devices for rapid NIR spectroscopy analysis to identify food fraud and ensure consumer safety. [ABSTRACT FROM AUTHOR]

The major histocompatibility complex (MHC) consists of genes involved in immune response and molecular discrimination between self and non-self. MHC genes are the most polymorphic in vertebrates. The origin and maintenance of polymorphism in MHC genes in populations is still unresolved. Mechanisms such as sexual selection and heterozygote advantage have been suggested as explanations for this high variability. In this study, a farmed population of rainbow trout (Oncorhynchus mykiss) characterized by the presence of specific MHC class IIB gene haplotypes at a frequency higher (30%) than that expected from random matings was investigated. Therefore, it was hypothesized that disassortative matings occur with an adaptive advantage for females, resulting in improved reproductive performance when mated with individuals with similar MHC haplotypes. Genetic analyses of the breeders were performed to define the MHC haplotypes and to perform specific matings. The effect of mating was evaluated by analyzing the survival rate of the offspring at various stages of incubation until swim-up. The reproductive performance of the offspring derived from specimens with similar haplotypes showed a better survival trend during the first life stages and reduced malformations. The results obtained are in contrast with the heterozygous advantage theory, therefore it was hypothesized, as for other salmonid species, the presence of a positive selection towards locally adapted MHC genes that promotes reproduction between genetically similar individuals. [ABSTRACT FROM AUTHOR]

Simple Summary: Reptiles usually carry Salmonella without showing any signs of infection. In this case, an adult male corn snake died 48 h after eating a feeder mouse bought online. The snake and mouse livers tested positive for Salmonella, specifically Salmonella enterica subsp. enterica serovar Midway. A genome analysis revealed that the two samples were from the same bacterial strain, and both had genes responsible for the bacteria's virulence. This report is about a pet snake that acquired Salmonella from a feeder mouse and then died from septic shock. It shows how feeder mice can be a source of clinical salmonellosis in pet snakes, which can pose a risk to humans too. Reptiles are usually asymptomatic carriers of Salmonella, with the manifestation of typical clinical signs of acute forms in adult and non-immunocompromised animals being considered exceptions. In the present case, an adult male corn snake (Pantherophis guttatus) was found dead due to septic shock 48 h after consuming a feeder mouse purchased online. The snake's tissue samples and faeces were cultured for bacteria isolation. Microbiological examinations of the snake and mouse livers revealed the presence of Salmonella enterica subsp. enterica serovar Midway. A whole-genome analysis of these two isolates showed a high correlation between them: they belonged to the strain type ST-357 for the classic MLST scheme and to the strain type ST 171322 for the cgMLST scheme. Also, a virulence gene analysis revealed the presence of stdB and STM3026 genes. This report conveys a case of food-borne salmonellosis in a pet snake, transmitted from a feeder mouse, likely responsible for the snake's death due to septic shock. It highlights the relevance of feeder mice as a source of Salmonella infections in snakes and the associated risks to human health. [ABSTRACT FROM AUTHOR]

High-mountain lakes are freshwater ecosystems situated above the tree line which are known for their remote locations and limited accessibility. These ecosystems host simplified biotic communities primarily concentrated in the littoral zone and dominated by benthic macroinvertebrates that serve as bioindicators of environmental pressures. A two-year monitoring investigation was performed in July 2022 and July 2023 at Nero Lake (Cesana Torinese, Northwest Italy). Five sites along the lakeshore were selected for sampling physicochemical water parameters and macrobenthos. All collected data were analysed to compare trends across years and within specific sites. The results revealed that Nero Lake exhibited consistent macrobenthic communities across the two years studied, but significant differences were observed in its microhabitats. This suggests that substrate type and physicochemical water parameters strongly influence community composition. Chironomidae larvae and Mollusca were the dominant species, showing distinct associations with different substrates and environmental factors from one year to another. These findings contribute to our understanding of the intricate relationships between benthic macroinvertebrates and their environments, highlighting the necessity of detailed, small-scale assessments to comprehend ecosystem dynamics and develop effective conservation strategies. [ABSTRACT FROM AUTHOR]

Simple Summary: Both wild (Sorghum halepense) and cultivated Sorghum (Sorghum bicolor) species are commonly used for animal feeding. However, sorghum plants naturally produce dhurrin, a cyanogenic glycoside releasing cyanide; this extremely toxic molecule potentially causes lethal toxicoses, particularly in ruminants. This was the case in a number of farms located in Northwest Italy during August 2022, under weather conditions (drought, tropical temperatures) known to increase plants' dhurrin content. Sixty-six bovines died after grazing Sorghum pastures (Sorghum bicolor or Sorghum halepense) or being fed with Sorghum-containing hay (Sorghum halepense). The reported clinico-pathological findings clearly indicated cyanide poisoning, and chemical analysis revealed high concentrations of dhurrin in the plant materials. The successful management of such toxicosis should rely on the prompt removal of the contaminated fodder and the administration of the antidote sodium thiosulphate. Dhurrin content should be carefully monitored, particularly in the hot season, and both farmers and veterinarians should be made aware of the risks associated with feeding cattle even with cultivated Sorghum, particularly if grown under adverse climatic conditions. Sorghum plants naturally produce dhurrin, a cyanogenic glycoside that may be hydrolysed to cyanide, resulting in often-lethal toxicoses. Ruminants are particularly sensitive to cyanogenic glycosides due to the active role of rumen microbiota in dhurrin hydrolysis. This work provides an overview of a poisoning outbreak that occurred in 5 farms in Northwest Italy in August 2022; a total of 66 cows died, and many others developed acute toxicosis after being fed on either cultivated (Sorghum bicolor) or wild Sorghum (Sorghum halepense). Clinical signs were recorded, and all cows received antidotal/supportive therapy. Dead animals were subjected to necropsy, and dhurrin content was determined in Sorghum specimens using an LC–MS/MS method. Rapid onset, severe respiratory distress, recumbency and convulsions were the main clinical features; bright red blood, a bitter almond smell and lung emphysema were consistently observed on necropsy. The combined i.v. and oral administration of sodium thiosulphate resulted in a rapid improvement of clinical signs. Dhurrin concentrations corresponding to cyanide levels higher than the tolerated threshold of 200 mg/kg were detected in sorghum specimens from 4 out of 5 involved farms; thereafter, such levels declined, reaching tolerable concentrations in September–October. Feeding cattle with wild or cultivated Sorghum as green fodder is a common practice in Northern Italy, especially in summer. However, care should be taken in case of adverse climatic conditions, such as severe drought and tropical temperatures (characterising summer 2022), which are reported to increase dhurrin synthesis and storage. [ABSTRACT FROM AUTHOR]

Simple Summary: Testosterone and nandrolone can be illegally administered to meat-producing animals as synthetic esters. To tackle the abuse of growth promoters, alternative approaches able to investigate specific changes induced in proteins, transcripts, and metabolites are becoming recommendable. This work aimed to characterize transcriptome perturbations related to the illicit administration of steroid esters in fattening pigs. Animals were treated with testosterone esters (Sustanon®, Organon, Jersey City, NJ, USA) or nandrolone esters (Myodine®, Le Vet Beheer B.V., Oudewater, Utrecht, The Netherlands). At the end of the trial, liver samples were collected for gene expression studies. Comparisons between treated and control groups using RNAseq allowed the identification of 491 differentially expressed genes (DEGs). Further analysis of DEGs characterized a smaller cluster of 16 genes. A field survey performed on liver samples collected from pigs belonging to different breeds and weight categories allowed the validation of the selected biomarkers using qPCR, confirming their specificity by comparison with testosterone residue profiles on respective serum samples. The use of anabolic–androgenic steroids (AASs) as growth promoters in farm animals is banned in the European Union, representing both an illicit practice and a risk for consumer health. However, these compounds are still illegally administered, often in the form of synthetic esters. This work aimed to characterize significant coding RNA perturbations related to the illicit administration of testosterone and nandrolone esters in fattening pigs. A total of 27 clinically healthy 90-day-old pigs were randomly assigned to test and control groups. Nine animals were treated with testosterone esters (Sustanon®) and other nine with nandrolone esters (Myodine®). At the end of the trial, liver samples were collected and analyzed using RNAseq, allowing the identification of 491 differentially expressed genes (DEGs). The transcriptional signature was further characterized by a smaller sub-cluster of 143 DEGs, from which a selection of 16 genes was made. The qPCR analysis confirmed that the identified cluster could still give good discrimination between untreated gilt and barrows compared to the relative testosterone-treated counterparts. A conclusive field survey on 67 liver samples collected from pigs of different breeds and weight categories confirmed, in agreement with testosterone residue profiles, the specificity of selected transcriptional biomarkers, showing their potential applications for screening purposes when AAS treatment is suspected, allowing to focus further investigations of competent authorities and confirmatory analysis where needed. [ABSTRACT FROM AUTHOR]

Lactococcus petauri is a recently described species of the genus Lactococcus. It was reported as an etiological agent of piscine lactococcosis together with Lactococcus garvieae. L. garvieae was already described as an opportunistic pathogen in human infections, with a potential zoonotic role. This paper represents the first report of a human urinary tract infection caused by L. petauri. A 91-year-old man was admitted to the emergency department for a femur fracture consequent to a domestic accident. The fracture was reduced by surgery and a catheterized specimen urine culture revealed a high bacterial load sustained by Gram-positive cocci, identified by Vitek 2 compact as L. garvieae, and subsequently as L. petauri through Internal Transcribed spacer 16S-23S r-RNA amplification. The number of L. petauri infections in humans is expected to rise in the near future mainly due to diagnostic improvement. A dedicated survey on L. garvieae and L. petauri infections in humans should be performed to better understand their role as pathogens and as zoonotic agents. [ABSTRACT FROM AUTHOR]

The gut microbiota has become a topic of increasing importance in various fields, including aquaculture. Several fish species have been the subject of investigations concerning the intestinal microbiota, which have compared different variables, including the intestinal portions, the environment, and diet. In this study, the microbiota of farmed and wild brook trout (Salvelinus fontinalis) were analyzed, in which the wall and content of the medial portion of the intestine were considered separately. A total of 66 fish (age class 2+) were sampled, of which 46 were wild and 20 were farmed brook trout, in two different years. Microbiota data were obtained using a 16S metabarcoding approach by analyzing the V3–V4 hypervariable regions of the corresponding 16S rRNA. The data showed that the core microbiota of these species consist of Proteobacteria (Alpha- and Gammaproteobacteria), Actinobacteria, Firmicutes (Bacilli and Clostridia), and, only for farmed animals, Fusobacteria. The latter taxon's presence is likely related to the fishmeal-based diet administered to farmed brook trout. Indeed, alpha and beta diversity analysis showed differences between wild and farmed fish. Finally, statistically significant differences in the microbiota composition were observed between the intestinal walls and contents of wild fish, while no differences were detected in reared animals. Our work represents the first study on the intestinal microbiota of brook trout with respect to both farmed and wild specimens. Future studies might focus on the comparison of our data with those pertaining to other fish species and on the study of other portions of the brook trout intestine. [ABSTRACT FROM AUTHOR]

Among forbidden substances included in the European Union legislation, endogenous steroids constitute a challenge in the framework of veterinary Official Monitoring Plans. They can be naturally present in body fluids at variable levels depending on the species, sex and age of the animals. Considering the significant advances achieved in breeding conditions and in the selection of producing traits in meat cattle, the aim of this study was to verify by analytical method, in veal calves housed under controlled conditions, if the level of natural steroids hormones assumed as physiological are still actual. The second aim of the study was to verify if the normal histological pattern of growth promoters in target organs is influenced accordingly. Bovine male sex organs are currently analysed in the frame of the Italian histological plan to monitor illicit treatments trend, highlighting microscopic, induced alterations. The levels of 17β-estradiol and progesterone residues resulted under the Limit of Quantitation of the approved official methods and the level of testosterone resulted below the level stated in the Italian Ministerial Decree in force. Male target organs appeared within the limits of the standard histological features. [ABSTRACT FROM AUTHOR]

Swinepox virus (SWPV) is responsible for sporadic acute poxvirus infections in swine worldwide, causing a pathognomonic eruptive proliferative dermatitis. Beside direct and congenital transmission, the pig louse Haematopinus suis acts as a mechanical vector and favors virus infection through skin lesions. Infections are generally described in domestic pigs, while only a few cases have been reported in wild boars, in Austria and Germany. In September 2022, SWPV infection was suspected at post-mortem examination of a wild boar piglet with characteristic lesions in Liguria, Northwest Italy. The piglet was heavily parasitized by swine lice (H. suis). SWPV was then confirmed by histological and molecular analyses. Possible viral co-infections were also investigated (African swine fever virus, classical swine fever virus, parvovirus, circovirus, Aujeszky's disease virus and hepatitis E virus). This article describes gross and histopathologic features of SWPV infection, differential diagnosis, and potential vector-borne transmission to domestic pigs, presenting a brief review of the literature on the topic. SWPV infection is reported in wild boars in Italy for the first time. The finding of SWPV in a wild boar in an area with a very limited pig population may suggest the existence of a "wildlife cycle" in the area. Further investigations are needed to understand the real risk of transmission of SWPV to domestic pigs as well as the role of other arthropod vectors. [ABSTRACT FROM AUTHOR]

Plastic is a polymer extremely resistant to degradation that can remain for up to hundreds or thousands of years, leading to the accumulation of massive amounts of plastic waste throughout the planet's ecosystems. Due to exposure to various environmental factors, plastic breaks down into smaller particles named microplastics (1–5000 μm) and nanoplastics (<1 μm). Microplastics (MPs) are ubiquitous pollutants but, still, little is known about their effects on human and animal health. Herein, our aim is to investigate cytotoxicity, oxidative stress, inflammation and correlated gene modulation following exposure to polystyrene microplastics (PS-MPs) in HRT-18 and CMT-93 epithelial cell lines. After 6, 24 and 48 h PS-MPs treatment, cell viability (MTT) and oxidative stress (SOD) assays were performed; subsequently, expression changes and cytokines release were investigated by Real-Time PCR and Magnetic-beads panel Multiplex Assay, respectively. For each exposure time, a significantly increased cytotoxicity was observed in both cell lines, whereas SOD activity increased only in CMT-93 cells. Furthermore, Magnetic-beads Multiplex Assay revealed an increased release of IL-8 in HRT-18 cells' medium, also confirmed by gene expression analysis. Results obtained suggest the presence of a pro-inflammatory pattern induced by PS-MPs treatment that could be related to the observed increase in cytotoxicity. [ABSTRACT FROM AUTHOR]

Simple Summary: PIVKA-II is an aberrant form of vitamin K that is increased in human coagulation disfunctions and in some neoplastic diseases. In veterinary medicine, PIVKA-II concentrations can be useful to identify patients with coagulative disorders in plasma and tissues, but its role as marker for hepatocellular carcinoma has not been investigated previously. In this study we characterized ed the expression of PIVKA-II in canine hepatocellular carcinomas in relation with the prognosis and some histological grading systems with the aim of finding useful prognostic factors for this canine tumour. Background: Hepatocellular carcinoma (HCC) in dogs is uncommon and often associated with a good prognosis, although some cases prove to be aggressive. In human oncology HCC is often very aggressive and diagnostic methods and prognostic factors are widely used to predict its biological behaviour. These include the expression of PIVKA-II. Methods: in order to identify a prognostic factor for canine HCC, we applied different methods of histological grading and investigated PIVKA-II expression in 22 HCC of dogs treated surgically and followed clinically for at least 2 years. Results: Nineteen patients analysed have passed the observation period without tumour recurrence, while 3 died following the development of metastases. PIVKA-II was positive in 15/22 cases without correlation with prognosis or tumoural grading even if a trend of PIVKA-II negativity in low WHO grades as well as increased number of PIVKA-II positive cases in higher WHO grades weres observed. Conclusions: This work showed that, PIVKA-II cannot be considered either as a marker of malignancy or as a prognostic marker for canine HCC. The poor prognosis depends usually on the clinical presentation. Thus prognostic parameters in canine HCC able to predict its aggressive behaviour through histological examination are still missing. The most promising method, limited to our study, seems to be the WHO histological grading. [ABSTRACT FROM AUTHOR]

Simple Summary: Canine mammary cancer is very common and has many similarities with human breast cancer. Risk factors, physiological and pathological behaviors, and the clinical course in dogs are very similar to humans. Several molecular similarities have also been reported, such as overexpression of EGF, proliferation markers, metalloproteinase and cyclooxygenase, TP53 mutations, and CXCR4/SDF1 axis activation. These common characteristics make these breast tumors resistant to conventional therapies. It is therefore necessary to study therapeutic alternatives. Cell lines could be helpful to test in vitro immunomodulant anti-cancer therapies, allowing a reduction of laboratory animals' involvement in the preliminary tests and achieving results in a shorter time. Although the canine mammary carcinoma cell line CF33 has been widely used in many studies on dog mammary cancer, characterization of its gene expression profile and of the influence of infective stressors of this cell line is poor. Our study shows the interaction of CF33 and Salmonella Typhimurium (ST) as an infective stressor, indicating that these cells may represent an in vitro model for assessing novel therapeutic approaches using bacteria. Spontaneous mammary tumors are the most frequent neoplasms in bitches and show similarities with human breast cancer in risk factors, clinical course, and histopathology. The poor prognosis of some cancer subtypes, both in human and dog, demands more effective therapeutic approaches. A possible strategy is the new anticancer therapy based on immune response modulation through bacteria or their derivatives on canine mammary carcinoma cell lines. The aim of the present study was to analyze the CF33 cell line in terms of basal expression of immune innate genes, CXCR4 expression, and interaction with infectious stressors. Our results highlight that CF33 maintains gene expression parameters typical of mammary cancer, and provides the basal gene expression of CF33, which is characterized by overexpression of CXCR4, CD44, RAD51, LY96, and a non-continuous expression of TP53 and PTEN. No mutations appeared in the CXCR4 gene until the 58th passage; this may represent important information for studying the CXCR4 pathway as a therapeutic target. Moreover, the CF33 cell line was shown to be able to interact with Salmonella Typhimurium (ST) (an infective stressor), indicating that these cells could be used as an in vitro model for developing innovative therapeutic approaches involving bacteria. [ABSTRACT FROM AUTHOR]

Simple Summary: Advances in cancer research are crucial, and pet oncology can improve the knowledge in several ways. Dogs are not only models of specific naturally occurring tumours but can also be sentinels of environmental exposures to carcinogenic substances, as they share the same environment with their owners. The purpose of this work was to describe the data collected by The Italian Network of Laboratories for Veterinary Oncology in the first 9 years of activity (2013–2021) and to evaluate their potential epidemiological significance. Frequencies of tumour sites in dogs were described, analysed and compared, considering several risk factors (breed, sex, period and region of residence). These observations allowed us to highlight differences not only in the site of occurrence of some tumours but also to formulate hypotheses on the potential role of some risk factors, e.g., neutering/spaying or geographical location. In our opinion, the results of this study confirm the importance of initiating and consolidating animal cancer registration initiatives that would facilitate the possibility of conducting multicentric collaborative studies to deepen the knowledge of the epidemiology of tumours in dogs and, from a comparative perspective, in humans. Advances in tumour research are crucial, and comparative oncology can improve the knowledge in several ways. Dogs are not only models of specific naturally occurring tumours but can also be sentinels of environmental exposures to carcinogens, as they share the same environment with their owners. The purpose of this work was to describe the data collected by The Italian Network of Laboratories for Veterinary Oncology in the first 9 years of activity (2013–2021) and to evaluate their potential epidemiological significance. Frequencies of tumour topographies and main morphologies in dogs were described, analysed and compared, calculating age-adjusted proportional morbidity ratios and considering several risk factors (breed, sex, period and region of residence). These observations allowed us to highlight differences not only in morphology and topography of some tumours but also to formulate hypotheses on the potential role of some risk factors, e.g., neutering/spaying or geographical location. In our opinion, the results of this case series confirm the importance of initiating and consolidating animal cancer registration initiatives that would facilitate the possibility of conducting multicentric collaborative studies to deepen the knowledge of the epidemiology of tumours in dogs from a comparative perspective. [ABSTRACT FROM AUTHOR]

Corticosteroids such as Dexamethasone (DEX) are commonly licensed for therapy in meat animals due to their known pharmacological properties. However, their misuse aimed to achieve anabolic effects is often found by National Residues Control Plans. The setup of a complementary "biomarker based" methods to unveil such illicit practices is encouraged by current European legislation. In this study, the combined use of molecular and histological quantitative techniques was applied on formalin fixed paraffin embedded (FFPE) muscle samples to assess the effects of illicit DEX treatment on veal calves. A PCR array, including 28 transcriptional biomarkers related to DEX exposure, was combined with a histochemical analysis of muscle fiber. An analysis based on unsupervised (PCA) and supervised (PLS-DA and Kohonen's SOM) methods, was applied in order to define multivariate models able to classify animals suspected of illicit treatment by DEX. According to the conventional univariate approach, a not-significant reduction in type I fibres was recorded in the DEX-treated group, and only 12 out of 28 targeted genes maintained their expected differential expression, confirming the technical limitations of a quantitative analysis on FFPE samples. However, the multivariate models developed highlighted the possibility to establish complementary screening strategies, particularly when based on transcriptional biomarkers characterised by low expression profiles. [ABSTRACT FROM AUTHOR]

European sea bass (Dicentrarchus labrax L.) is one of the most economically important fish species in the Mediterranean Sea area. Despite strict requirements regarding indications of production method (wild/farmed), incorrect labelling of sea bass is a practice still frequently detected. The aim of this study was to evaluate the capabilities of two techniques, Near-InfraRed (NIR) spectroscopy and mass spectrometry, to discriminate sea bass according to the production method. Two categories were discriminated based on the docosahexaenoic and arachidonic fatty acid ratio by using a Direct Sample Analysis (DSA) system integrated with a time-of-flight (TOF) mass spectrometer. The cut-off value of 3.42, of fatty acid ratio, was able to discriminate between the two types of fish with sensitivity and specificity of 100%. It was possible to classify fish production by using multivariate analysis with portable NIR. The results achieved by the developed validation models suggest that this approach is able to distinguish the two product categories with high sensitivity (100%) and specificity (90%). The results obtained from this study highlight the potential application of two easy, fast, and accurate screening methods to detect fraud in commercial sea bass production. [ABSTRACT FROM AUTHOR]

Fish species substitution is one of the most common forms of fraud all over the world, as fish identification can be very challenging for both consumers and experienced inspectors in the case of fish sold as fillets. The difficulties in distinguishing among different species may generate a "grey area" in which mislabelling can occur. Thus, the development of fast and reliable tools able to detect such frauds in the field is of crucial importance. In this study, we focused on the distinction between two flatfish species largely available on the market, namely the Guinean sole (Synaptura cadenati) and European plaice (Pleuronectes platessa), which are very similar looking. Fifty fillets of each species were analysed using three near-infrared (NIR) instruments: the handheld SCiO (Consumer Physics), the portable MicroNIR (VIAVI), and the benchtop MPA (Bruker). PLS-DA classification models were built using the spectral datasets, and all three instruments provided very good results, showing high accuracy: 94.1% for the SCiO and MicroNIR portable instruments, and 90.1% for the MPA benchtop spectrometer. The good classification results of the approach combining NIR spectroscopy, and simple chemometric classification methods suggest great applicability directly in the context of real-world marketplaces, as well as in official control plans. [ABSTRACT FROM AUTHOR]

Chronic wasting disease (CWD) is a prion disease that affects cervids; it is classified under transmissible spongiform encephalopathies (TSEs). CWD is particularly contagious, making its eradication in endemic areas very difficult and creating serious problems for cervid conservation and breeding. It has recently become an emerging public health risk to be managed by health authorities. Starting in 2017, active CWD surveillance in Italy has intensified with the monitoring of wild and farmed cervids. The present study summarizes findings from a histopathological survey of the brains from wild ruminants collected via CWD monitoring between 2017 and 2019. A total of 113 brains from 62 red deer (Cervus elaphus) and 51 roe deer (Capreolus capreolus) were submitted for analysis at the National Reference Center for Animal Encephalopathies (CEA) to determine major patterns of neuropathological lesions and correlated pathogens. Brain lesions were detected in 20 animals, 10 brain samples were unsuitable for examination, and 83 presented no lesions. Neuropathological examination revealed non-suppurative encephalitis or meningoencephalitis in most cases (15/20). This brain study revealed evidence for the absence of CWD in Italy and provided a reference spectrum of neuropathological lesions for differential diagnosis in cervids. [ABSTRACT FROM AUTHOR]

Although no mass mortality has been recorded so far, the precise demographic effect of white-nose syndrome (WNS) on European bats still remains to be ascertained. Following the first isolation of P. destructans in Italy, further surveys were performed to assess the distribution of the fungus in NW Italy and its effects on bats. Data were collected from March 2019 to April 2020 at sites used for hibernation (six sites) and/or for reproduction (four sites) in Piedmont and Aosta Valley. A total of 138 bats, belonging to 10 species, were examined to identify clinical features possibly related to the fungal presence. Culture from swabs and the molecular identification of isolates confirmed the presence of P. destructans in bats from five sites, including two maternal roosts. Dermal fungal infiltration, the criterion to assess the presence of WNS, was observed in biopsies of bats belonging to Myotis blythii, M. daubentonii, M. emarginatus and M. myotis. This is the first report of the disease in Italy. The results suggest a greater susceptibility to the infection of the genus Myotis and particularly of M. emarginatus, possibly due to the long length of its hibernation period. Other fungal dermatophytes were also observed. [ABSTRACT FROM AUTHOR]

Recombinant bovine growth hormone (rbGH) is produced in large quantities and widely used in a number of countries worldwide to stimulate milk production in dairy animals. The use of this compound in animal production is strictly regulated by food safety directives in force, in particular in the European Union (EU). Although analytical strategies for the detection of rbGH in blood have been successfully reported over the past 15 years, they do not fully answer the expectations of either competent authorities or industrials that would expect measuring its occurrence directly in the milk. As a matrix of excretion but also of consumption, milk appears indeed as the matrix of choice for detecting the use of rbGH in dairy animals. It also allows large volumes to be collected without presenting an invasive character for the animal. However, rbGH detection in milk presents several challenges, mainly related to the sensitivity required for its detection in a complex biological matrix. This review article presents the specific difficulties associated with milk and provides an overview of the analytical strategies reported in the literature and whether they concern indirect or direct approaches to the detection of rbGH administration to animals, with applications either for screening or confirmation purposes. [ABSTRACT FROM AUTHOR]

Simple Summary: PIVKA-II is an aberrant form of vitamin K that has been demonstrated to be increased in human coagulation disorders and in some neoplastic diseases. In veterinary medicine, PIVKA-II levels have been demonstrated to be useful for distinguishing anticoagulant poisoning from other coagulopathies. In forensic pathology, there is the need to distinguish malicious poisoning from other causes of death and, in some cases, identifying poisoned dogs from dogs that died as a result of other coagulative disorders can be challenging. This study evaluated the usefulness of the immunohistochemical expression of PIVKA-II in hepatic and renal tissues in order to identify patients with coagulative disorders due to clinical condition or the ingestion of anticoagulants substances. PIVKA-II is an aberrant form of vitamin K that has been demonstrated to be increased in human coagulation disorders and in some neoplastic diseases. In veterinary medicine, PIVKA-II levels have been demonstrated to be useful for distinguishing anticoagulant poisoning from other coagulopathies. In forensic pathology, there is the need to distinguish malicious poisoning from other causes of death and, in some cases, identifying poisoned dogs from dogs that died as a result of other coagulative disorders can be challenging. In this study, dogs that suddenly died underwent necropsy, histological examination, and toxicological analysis to establish cause of death. PIVKA-II immunohistochemical expression was evaluated on hepatic and renal tissues, and on neoplastic lesions when present. A total of 61 dogs were analyzed and anticoagulant substances were identified in 16 of the 61. Immunolabelling for PIVKA-II was observed in 27 of 61 cases in the liver and in 24 of 61 cases in the kidneys. Among the poisoned dogs, the PIVKA-II expression was present in the liver in 15 of 16 cases and in the kidneys in 16 of 16. Neoplastic lesions represented mainly by haemangiosarcomas were negative. This study highlights how the immunohistochemical expression of PIVKA-II in hepatic and renal tissues can be useful to identify patients with coagulative disorders due to clinical condition or the ingestion of anticoagulants substances. [ABSTRACT FROM AUTHOR]

Fish mycobacteriosis is a widespread global problem caused by species of non-tuberculous mycobacteria (NTM). Mycobacterium marinum is one of the species most often involved in disease episodes of aquarium and farmed fish. Since there is currently no available effective therapy or vaccine, a prompt search for routes of entry is key to limiting the damage induced by the disease. Here we report a case of mycobacteriosis follow up in a European sea bass (Dicentrarchus labrax) farm located in Northern Italy, in which environmental samples and newly added fish batches were analyzed. Samples from fish present on the farm, sediment, and periphyton all resulted positive for M. marinum, whereas the new fish batches and the water samples resulted negative. The environmental resistance of NTM (alcohol-acid resistance, biofilm formation) and the lack of prophylactic and therapeutic strategies make these diseases difficult to manage. Prompt identification of biotic and abiotic reservoirs, combined with good zootechnical hygiene practices, are the most effective measures to control fish mycobacteriosis in intensive farms. [ABSTRACT FROM AUTHOR]

Simple Summary: Growing concern exists that SARS-CoV-2, as has already been ascertained for its SARS-CoV and MERS-CoV "predecessors", originated from an animal "reservoir", thereafter spilling over into mankind, possibly anticipated by a viral "passage" into a secondary animal host. Within the dramatic SARS-CoV-2 pandemic context, hitherto characterized by over 110 million cases and almost 2,500,000 deaths on a global scale, several domestic and wild animal species have been reported as being susceptible to natural and/or experimental SARS-CoV-2 infection. In this respect, while some marine mammal species are deemed as potentially susceptible to SARS-CoV-2 infection on the basis of the sequence homology of their ACE-2 viral receptor with the human one, this study addresses this critical issue in stranded sea mammal specimens. Zoonotically transmitted coronaviruses were responsible for Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2), causing the dramatic Coronavirus Disease-2019 (CoViD-19) pandemic, which affected public health, the economy, and society on a global scale. The impact of the SARS-CoV-2 pandemic permeated into our environment and wildlife as well; in particular, concern has been raised about the viral occurrence and persistence in aquatic and marine ecosystems. The discharge of untreated wastewaters carrying infectious SARS-CoV-2 into natural water systems that are home to sea mammals may have dramatic consequences on vulnerable species. The efficient transmission of coronaviruses raises questions regarding the contributions of virus-receptor interactions. The main receptor of SARS-CoV-2 is Angiotensin Converting Enzyme-2 (ACE-2), serving as a functional receptor for the viral spike (S) protein. This study aimed, through the comparative analysis of the ACE-2 receptor with the human one, at assessing susceptibility to SARS-CoV-2 for different species of marine mammals living in Italian waters. We also determined, by means of immunohistochemistry, ACE-2 receptor localization in the lung tissue from different cetacean species, in order to provide a preliminary characterization of ACE-2 expression in the marine mammal respiratory tracts. Furthermore, to evaluate if and how Italian wastewater management and coastal exposition to extreme weather events may led to susceptible marine mammal populations being exposed to SARS-CoV-2, geomapping data were carried out and overlapped. The results showed the potential SARS-CoV-2 exposure for marine mammals inhabiting Italian coastal waters, putting them at risk when swimming and feeding in specific risk areas. Thus, we highlighted the potential hazard of the reverse zoonotic transmission of SARS-CoV-2 infection, along with its impact on marine mammals regularly inhabiting the Mediterranean Sea, while also stressing the need for appropriate action in order to prevent further damage to specific vulnerable populations. [ABSTRACT FROM AUTHOR]

The sale of frozen–thawed fish and fish products, labeled as fresh, is currently one of the most common and insidious commercial food frauds. For this reason, the demand of reliable tools to identify the storage conditions is increasing. The present study was performed on two species, commonly sold in large-scale distribution: Cuttlefish (Sepia officinalis) and musky octopus (Eledone spp.). Fifty fresh cephalopod specimens were analyzed at refrigeration temperature (2 ± 2 °C), then frozen at −20 °C for 10 days and finally thawed and analyzed again. The performance of three near-infrared (NIR) instruments in identifying storage conditions were compared: The benchtop NIR Multi Purpose Analyzer (MPA) by Bruker, the portable MicroNIR by VIAVI and the handheld NIR SCiO by Consumer Physics. All collected spectra were processed and analyzed with chemometric methods. The SCiO data were also analyzed using the analytical tools available in the online application provided by the manufacturer to evaluate its performance. NIR spectroscopy, coupled with chemometrics, allowed discriminating between fresh and thawed samples with high accuracy: Cuttlefish between 82.3–94.1%, musky octopus between 91.2–97.1%, global model between 86.8–95.6%. Results show how food frauds could be detected directly in the marketplace, through small, ultra-fast and simplified handheld devices, whereas official control laboratories could use benchtop analytical instruments, coupled with chemometric approaches, to develop accurate and validated methods, suitable for regulatory purposes. [ABSTRACT FROM AUTHOR]

Mycobacterium pseudoshottsii, a slow-growing mycobacterium closely related to M. marinum, has been isolated only in wild fish in the United States and in Japanese fish farms to date. Here, we report cases of mortality in three farmed fish species (Dicentrarchus labrax, Sparus aurata, and Sciaenops ocellatus) caused by M. pseudoshottsii in Italy. Samples underwent necropsy, histology, and culture with pathogen identification based on PCR and sequencing of housekeeping genes (16S rRNA, hsp65, rpoB). Multifocal to coalescing granulomatous and necrotizing inflammation with acid-fast bacilli were observed in the parenchymatous organs, from which M. pseudoshottsii was isolated and identified. Phylogenetic analysis confirmed the results of gene sequencing and allowed subdivision of the isolates into three distinct groups. M. pseudoshottsii poses a potential threat for Mediterranean aquaculture. Its origin in the area under study needs to be clarified, as well as the threat to the farmed fish species. [ABSTRACT FROM AUTHOR]

Shellfish farming is a relevant economic activity in Italy. The Gulf of La Spezia is one of the major production areas for mussels: the area is characterized by the presence of numerous human activities that could harm the quality of seawater. Additionally, the presence of potentially pathogenic microorganisms may influence the health status of animals, which must be constantly monitored. To have a clear view of the health conditions of the mussels (Mytilus galloprovincialis) farmed in this area, microbiological, parasitological, and histological analyses were performed. The study was conducted from November 2016 to October 2017. Overall, despite the presence of potentially pathogenic microorganisms for mussels, abnormal mortality rates were not reported during the monitoring period and the histological examination revealed no significant lesions. Our study confirms that studying different aspects together is a useful tool for assessing the health conditions of mussels and points out the importance of adverse environmental conditions for the expression of the pathogenicity of microorganisms. [ABSTRACT FROM AUTHOR]

The aim of this study was to describe liver alterations observed in a bullhead (Cottus gobio) population from a high-mountain lake (Dimon Lake; 1857 m a.s.l.) located in Carnic Alps (Northeast Italy). Two fish sampling campaigns (summer and autumn) were performed in 2017 to explore the possible causes of this phenomenon. In addition, to investigate the typical liver features of C. gobio, control specimens were captured from Degano Creek in the same seasons. Total length, weight, and liver weight were recorded in fish from both sampling sites. In addition, Fulton's condition factor (K) and hepatosomatic index (HSI) were calculated. Stomach contents were also analyzed. Liver from each specimen was sampled, and histological examination was carried out. Liver steatosis (L) (nuclear displacement and cytoplasm vacuolization) were histologically evaluated by assigning a semiquantitative severity score. A significant difference in L was found between Dimon Lake and Degano Creek in both summer (Mann–Whitney test; p = 0.0001) and autumn (Mann–Whitney test; p < 0.0001). Regarding HSI, a significant difference was also recorded between Dimon Lake and Degano Creek in summer (Mann–Whitney test; p < 0.0001) and also in autumn (Mann–Whitney test; p < 0.0001), but no seasonal change in K values was recorded between the two sites in both summer (Mann–Whitney test; p = 0.8589) and autumn (Mann–Whitney test; p = 0.6415). A significant positive correlation between HSI and L was found (ρS 0.573). The causes of this abnormality might be related to adaptation by the fish to the high-altitude environment, accumulating lipids in the liver to tackle the adverse season, which lasts for 6 months of the year. Nonetheless, our study does not preclude steatosis induced by a nearly exclusive chironomids diet in bullhead from Dimon Lake. [ABSTRACT FROM AUTHOR]