Effects of Disinfectants on Bacterium Paenibacillus larvae in Laboratory Conditions.

American foulbrood is an infectious disease of the honeybee brood that causes multiple types of damage to beekeeping. The causative agent of the disease is the bacterium Paenibacillus larvae, which forms resistant infective spores and is viable for decades. After the eradication measures have been implemented, in cases of clinically visible disease, it is necessary to conduct effective final disinfections of equipment and tools. This study aimed to determine the effect of ten commercially available and commonly used disinfectants on certified strains of P. larvae under laboratory conditions, as well as to compare the obtained results among individual genotypes of P. larvae. Selected products were tested by determining the zone of inhibition using an agar diffusion test, a suspension test for viable bacteria, a surface disinfectant test, and a sporicidal effect in the suspension test. Incidin OxyFoam S and Sekusept Aktiv are both effective against all examined genotypes of P. larvae. Despadac and Despadac Secure have a bactericidal effect, but their sporocidal effect is not as satisfactory as that of Genox. Genoll does not exhibit a sporicidal effect, and Ecocide S at 1%, Bee protect H forte, and Bee protect F did not exhibit a satisfactory sporocidal effect. Additionally, EM® PROBIOTIC FOR BEES did not exhibit any bactericidal effect. The effective application of control measures and proper application of final disinfection can reduce the reoccurrence of visible clinical signs of disease, whereas methods of early diagnosis can significantly reduce the incidence of the disease.

How to Measure Human-Dog Interaction in Dog Assisted Interventions? A Scoping Review.

Human-dog interaction is the working tool through which the therapeutic, educational and recreational goals of dog-assisted interventions (DAIs) are achieved. A better understanding of the characteristics of this interaction could improve the effectiveness of DAIs. This scoping review addresses the question: how has the human-dog connection been measured in the context of DAIs? After searching the Web of Science and Scopus platforms, only peer-reviewed, primary research studies reporting measures of therapy dog-human interaction, relationship and bond were included. A total of 70 included articles provided information on what was measured (interaction, relationship or bond) and how, as well as the general context (DAIs or experimental situations with therapy dogs). While the majority of the articles identified use behavioural analysis methods to analyse the interaction between the participant/recipient and the therapy dog during DAIs, it was possible to identify some more structured tools that assess the participant/recipient's interaction, relationship or bond with the therapy dog, as well as tools that consider the animal's perspective or focus on the dog-handler dyad, indicating growing areas of research. The tools and methods identified can be used by both practitioners and researchers to further explore aspects of human-dog interaction in the field of DAIs.

The oriental hornet (Vespa orientalis) as a potential vector of honey bee's pathogens and a threat for public health in North-East Italy.

BACKGROUND: Oriental hornets are large predatory hymenoptera that occur in the southern part of Asia and the southeastern Mediterranean. Among many pests of bee colonies, Vespa orientalis was recorded to be one of the most destructive. OBJECTIVES: The aim of this study was to: (1) monitor the presence of pathogens carried by V. orientalis that could potentially threaten honey bees and public health; (2) describe the hornet's predatory behavior on honey bee colonies and (3) collect the medical history of a V. orientalis sting suffered by a 36-year-old woman. METHODS: Observations of V. orientalis predatory behavior and the catches of hornets for parasitological and microbiological examination, using molecular and bacteriological analyses, were carried out in three experimental apiaries, both in spring in order to capture the foundress queens and during the summer to capture the workers. Furthermore, the medical history and photographic documentation of a V. orientalis sting suffered by a 36-year-old woman have been collected. RESULTS: The results obtained highlight that V. orientalis is capable of causing serious damage to beekeeping by killing bees, putting under stress the honey bee colonies and by potentially spreading honey bee pathogens among apiaries. These hornets may also become a public health concern, since they are capable of inflicting multiple, painful stings on humans. CONCLUSIONS: Only the development of an Integrated Management Control Program will be able to contain the negative effects of anomalous population growth and the potentially negative impact on honey bees and public health of V. orientalis.

A Pilot Study on Behavioural and Physiological Indicators of Emotions in Donkeys.

Recognizing animal emotions is critical to their welfare and can lead to a better relationship with humans and the environment, especially in a widespread species like the donkey, which is often prone to welfare issues. This study aims to assess the emotional response of donkeys through an operant conditioning task with two presumed different emotional contents. Specifically, a within-subject design including positive and negative conditions was conducted, collecting behavioural and physiological (heart rate variability and HRV) parameters. Facial expressions, postures, and movements were analysed by principal component analysis and behavioural diversity indexes (frequencies, activity budgets, richness, Shannon and Gini-Simpson). During the positive condition, both ears were held high and sideways (left: r = -0.793, p < 0.0001; right: r = -0.585, p = 0.011), while the ears were frontally erected (left: r = 0.924, p < 0.0001; right: r = 0.946, p < 0.0001) during the negative one. The latter was also associated with an increased tendency to walk (r = 0.709, p = 0.001), walk away (r = 0.578, p = 0.012), more frequent changes in the body position (VBody position = 0, p = 0.022), and greater behavioural complexity (VGini-Simpson Index = 4, p = 0.027). As for HRV analysis, the root mean square of successive beat-to-beat differences (rMSSD) was significantly lower after the negative condition. These non-invasive parameters could be considered as possible indicators of donkeys' emotional state.

Insects and Public Health: An Overview.

Insects are, by far, the most common animals on our planet. The ubiquity and plethora of ecological niches occupied by insects, along with the strict and sometimes forced coexistence between insects and humans, make insects a target of public health interest. This article reports the negative aspects historically linked to insects as pests and vectors of diseases, and describes their potential as bioindicators of environmental pollution, and their use as food and feed. Both negative and positive impacts of insects on human and animal health need to be addressed by public health professionals who should aim to strike a balance within the wide range of sometimes conflicting goals in insect management, such as regulating their production, exploiting their potential, protecting their health and limiting their negative impact on animals and humans. This requires increased insect knowledge and strategies to preserve human health and welfare. The aim of this paper is to provide an overview of traditional and emerging topics bridging insects and public health to highlight the need for professionals, to address these topics during their work. The present and future role and activities of public health authorities regarding insects are analyzed.

Donkey Heart Rate and Heart Rate Variability: A Scoping Review.

Heart rate (HR) and heart rate variability (HRV) are commonly used physiological measures in animals. While several studies exist on horse HRV, less information is available for donkeys. This scoping review aims to understand the extent and type of published evidence on donkey HR and HRV, their clinical and research applications, the devices used, and the analysis performed. Only quantitative primary studies published in English were considered. Four different databases were queried through the Web of Science platform, with additional evidence identified by citation chasing. After a two-stage screening phase, data were extracted considering study and population characteristics, information on HR/HRV analysis, and applications. The majority of the 87 included articles (about 80%) concerned a sample size of up to 20 individuals and were published since 2011 (about 65%). Forty-one articles employed an electronic device for signal acquisition (mainly electrocardiographs and heart rate monitors), yet only two articles reported HRV parameters. The literature on donkey HRV is lacking, and this gap can be filled by gaining knowledge on donkey characteristics and finding useful tools for welfare assessment. Comparison with what is known about the horse allows a discussion of the technical and interpretative difficulties that can be encountered with donkeys.

Some Considerations about Winter Colony Losses in Italy According to the Coloss Questionnaire.

The Italian beekeeping industry has grown steadily during the last decade, according to data from the national beekeeping registry, which came into existence in February 2015. Winter colony losses remain a matter of concern for beekeepers in Italy, and administration of the questionnaire defined by the Coloss Association could contribute to a better understanding of this phenomenon. To evaluate the percentage trends over time in honeybee colony losses arising from various causes, we used the quasi-binomial generalized linear modelling (GzLM) approach, taking the year as an independent variable. We set our level of significance at 5% and performed the data analysis only for the seven regions that sent data continuously from 2014 to 2020. We considered the percentage of losses due to queen-related problems, natural disasters, and dead or empty colonies, given that these questions remained unchanged over the years. The survey also revealed that the percentage trend for respondents using drone brood removal showed a significant increase. In general, the percentage of colony losses due to queen-related problems remained lower than 8%, and the percentage of colony losses associated with natural disasters was very low (<2%). The mean percentages of losses due to dead or empty colonies ranged from 6 to 17% in the considered period. In addition, we took account of the responses relating to treatments against Varroa mite infestation, given the importance attributed to this honeybee parasite. Unlike the other variables, we calculated the percentages related to the types of beekeeper treatments against Varroa destructor based on the respondents, not on the colonies. What emerged was that almost every beekeeper used at least one type of treatment against V. destructor. In general, the trend of respondents appeared stable at 0.3% during the last four years.

Disease control tools to secure animal and public health in a densely populated world.

Animal health is a prerequisite for global health, economic development, food security, food quality, and poverty reduction, while mitigating against climate change and biodiversity loss. We did a qualitative review of 53 infectious diseases in terrestrial animals with data from DISCONTOOLS, a specialist database and prioritisation model focusing on research gaps for improving infectious disease control in animals. Many diseases do not have any appropriate control tools, but the prioritisation model suggests that we should focus international efforts on Nipah virus infection, African swine fever, contagious bovine pleuropneumonia, peste des petits ruminants, sheeppox and goatpox, avian influenza, Rift Valley fever, foot and mouth disease, and bovine tuberculosis, for the greatest impact on the UN's Sustainable Development Goals. Easy to use and accurate diagnostics are available for many animal diseases. However, there is an urgent need for the development of stable and durable diagnostics that can differentiate infected animals from vaccinated animals, to exploit rapid technological advances, and to make diagnostics widely available and affordable. Veterinary vaccines are important for dealing with endemic, new, and emerging diseases. However, fundamental research is needed to improve the convenience of use and duration of immunity, and to establish performant marker vaccines. The largest gap in animal pharmaceuticals is the threat of pathogens developing resistance to available drugs, in particular for bacterial and parasitic (protozoal, helminth, and arthropod) pathogens. We propose and discuss five research priorities for animal health that will help to deliver a sustainable and healthy planet: vaccinology, antimicrobial resistance, climate mitigation and adaptation, digital health, and epidemic preparedness.

LC-HRMS-Based Non-Targeted Metabolomics for the Assessment of Honey Adulteration with Sugar Syrups: A Preliminary Study.

Honey is a natural product that is in great demand and has a relatively high price, thus making it one of the most common targets of economically motivated adulteration. Its adulteration can be obtained by adding cheaper honey or sugar syrups or by overfeeding honeybees with sugar syrups. Adulteration techniques are constantly evolving and advanced techniques and instruments are required for its detection. We used non-targeted metabolomics to underscore potential markers of honey adulteration with sugar syrups. The metabolomic profiles of unadulterated honeys and sugar beet, corn and wheat syrups were obtained using hydrophilic interaction liquid chromatography high-resolution mass spectrometry (LC-HRMS). The potential markers have been selected after data processing. Fortified honey (5%, 10% and 20%), honey obtained from overfeeding, and 58 commercial honeys were analyzed. One potential marker appeared with a specific signal for syrups and not for honey. This targeted analysis showed a linear trend in fortified honeys with a calculated limit of quantification around 5% of fortification.

Antimicrobial Activity from Putative Probiotic Lactic Acid Bacteria for the Biological Control of American and European Foulbrood Diseases.

The balance of the gut microbiome is important for the honey bee's growth and development, immune function and defense against pathogens. The use of a beneficial bacteria-based strategy for the prevention and biocontrol of American foulbrood (AFB) and European foulbrood (EFB) diseases in honey bees offers interesting prospects. Lactic acid bacteria (LAB) are common inhabitants of the gastrointestinal tract of the honey bee. Among LABs associated with bee gut microbiota, Lactiplantibacillus plantarum (previously Lactobacillus plantarum) and Apilactobacillus kunkeei (formerly classified as Lactobacillus kunkeei) are two of the most abundant species. In this study, four Lactiplantibacillus plantarum strains and four Apilactobacillus kunkeei strains, isolated from the gastrointestinal tract of honey bee (Apis mellifera L.) were selected for their in vitro inhibition ability of Paenibacillus larvae ATCC 9545 and Melissococccus plutonius ATCC 35311. In addition, these LABs have been characterized through some biochemical and functional characteristics: cell surface properties (hydrophobicity and auto-aggregation), carbohydrates assimilation and enzymatic activities. The antimicrobial, biochemical and cell surface properties of these LABs have been functional to their candidature as potential probiotics in beekeeping and for the biocontrol of AFB and EFB diseases.

A Systematic Review on Viruses in Mass-Reared Edible Insect Species.

Edible insects are expected to become an important nutrient source for animals and humans in the Western world in the near future. Only a few studies on viruses in edible insects with potential for industrial rearing have been published and concern only some edible insect species. Viral pathogens that can infect insects could be non-pathogenic, or pathogenic to the insects themselves, or to humans and animals. The objective of this systematic review is to provide an overview of the viruses detected in edible insects currently considered for use in food and/or feed in the European Union or appropriate for mass rearing, and to collect information on clinical symptoms in insects and on the vector role of insects themselves. Many different virus species have been detected in edible insect species showing promise for mass production systems. These viruses could be a risk for mass insect rearing systems causing acute high mortality, a drastic decline in growth in juvenile stages and in the reproductive performance of adults. Furthermore, some viruses could pose a risk to human and animal health where insects are used for food and feed.

Animal Assisted Interventions in the Green Care Framework: A Literature Review.

Green Care (GC) and Animal Assisted Interventions (AAI) are recognised practices useful to enhance the wellbeing of people through interaction with nature and animals. This study aims at understanding the interconnections between GC and AAI by analysing deeply which interaction with animals is conducted. Therefore, we carried out a literature search through Web of Science and Google Scholar that allowed retrieval of 993 records; after the PRISMA selection process, 42 were included. Relevant information was extracted: year of publication, geographical location, objectives, settings in agricultural environment, animal species, characteristics of users involved, type of human-animal interaction, coexistence of other activities without animals, animal health and welfare issues. From the review emerged that research on GC with animals is common in high-income countries and that the line between AAI and occupational therapy is often vague. Moreover, the most common setting for these interventions appears to be the farm, and frequently animals involved are not selected according to their ethological characteristics. Users in this context are extremely various and not only involved in activities with animals. Within the included studies, we noted a lack in the consideration of animal welfare that indicates the need for increased awareness among practitioners and a more ethical approach when animals are involved.

Effects of leukoreduction on storage lesions in whole blood and blood components of dogs.

BACKGROUND: Leukoreduction is a routine procedure in human transfusion medicine but is uncommon in veterinary. OBJECTIVES: To evaluate the effect of leukoreduction on the quality of canine whole blood (WB) and blood products during storage. ANIMALS: Ten canine blood donors. METHODS: This is a case series study. An amount of 450 mL of blood was collected from each dog. Five WB and 5 packed red blood cells (pRBC) bags were divided into 2 units each: leukoreduced (LR) and non-leukoreduced (nLR). RBC count, erythrocytes' mean osmotic fragility (MOF), 2,3-diphosphoglycerate (2,3-DPG), adenosine triphosphate (ATP), percentage of hemolysis, potassium (K), lactate, glucose, and cytokines were measured weekly from day of donation (T0) to day 35 (T35); pH, coagulation times, and clotting factors were evaluated at T0 and T35 from WB and in fresh frozen plasma after 1 year of storage. RESULTS: Leukoreduction showed positive effects on lactate (T35: LR WB 14.42 mmol/L SD 2.71, nLR WB 22.42 mmol/L SD 1.86, LR pRBC 20.88 mmol/L SD 2.65, nLR pRBC 36.81 mmol/L SD 2.34; P < .0001), pH (T35: LR WB 6.88 SD 0.16, nLR WB 6.69 SD 0.20, P = .02; LR pRBC 6.57 SD 0.23, nLR pRBC 6.22 SD 0.11; P < .001), and K (LR pRBC 4.08 mmol/L SD 0.88, nLR pRBC 5.48 mmol/L SD 0.90; P < .001). Increasing values of IL8 were observed in nLR units during storage (T0: 4167 ± 11 888 pg/mL; T35: 6367 ± 11 612 pg/mL). CONCLUSION AND CLINICAL IMPORTANCE: LR blood units are recommended to critically ill dogs with marked inflammatory conditions.

Antioxidant Activity in Bee Products: A Review.

Bee products have been used since ancient times both for their nutritional value and for a broad spectrum of therapeutic purposes. They are deemed to be a potential source of natural antioxidants that can counteract the effects of oxidative stress underlying the pathogenesis of many diseases. In view of the growing interest in using bioactive substances from natural sources to promote health and reduce the risk of developing certain illnesses, this review aims to update the current state of knowledge on the antioxidant capacity of bee products such as honey, pollen, propolis, beeswax, royal jelly and bee venom, and on the analytical methods used. The complex, variable composition of these products and the multitude of analytical methods used to study their antioxidant activities are responsible for the wide range of results reported by a plethora of available studies. This suggests the need to establish standardized methods to more efficiently evaluate the intrinsic antioxidant characteristics of these products and make the data obtained more comparable.

Genetic variability following selection for scrapie resistance in six autochthonous sheep breeds in the province of Bolzano (northern Italy).

Scrapie is an ovine transmissible spongiform encephalopathy, and its susceptibility is associated with polymorphisms in the prion protein gene (PRNP). Genetic selection is currently the most effective mean for eradication of the susceptible VRQ allele in favour of resistant ARR allele. Maintenance of genetic diversity should be one of the major objectives in breeding programmes, especially in endangered breeds, and genetic information are an excellent alternative to pedigree data where these information are missing. The aim of our study was to determine changes of genetic variability in six native sheep breeds from autonomous province of Bolzano, northern Italy, following simulation of scrapie selection scenarios. A total of 684 rams were investigated for PRNP polymorphisms and for 10 microsatellite loci to estimate genetic variability. Across all loci, a total of 163 alleles were detected with a mean of 10.4 alleles per locus. Average observed (Ho) and unbiased expected (uHe) heterozygosity overall loci were 0.74 and 0.78, respectively, showing a statistically significant deviation from Hardy-Weinberg equilibrium (HWE) in all breeds. This heterozygosity deficit was confirmed by a positive fixation index (Fis), determining a moderate inbreeding in each breed. Simulating a soft selection, where only rams having at least a VRQ allele should be excluded from reproduction, Ho, uHe and Fis values remained almost unchanged, indicating that genetic variability should not be affected by the removal of these individuals. With a mild selection scenario, considering only rams with at least one ARR allele, we observed a decrease in the mean alleles per breed (8.9) and the maintenance of heterozygosity deficiency, except for two breeds, where it was any longer significant. These results showed that selection strategies allowing use of heterozygous as well homozygous ARR rams might be the right compromise to improve resistance to scrapie and to do not dramatically affect genetic variability of these breeds.

Detection of replicative Kashmir Bee Virus and Black Queen Cell Virus in Asian hornet Vespa velutina (Lepelieter 1836) in Italy.

Information concerning the pathogenic role of honey bee viruses in invasive species are still scarce. The aim of this investigation was to assess the presence of several honey bee viruses, such as Black Queen Cell Virus (BQCV), Kashmir Bee Virus (KBV), Slow Paralysis Virus (SPV), Sac Brood Virus (SBV), Israeli Acute Paralysis Virus (IAPV), Acute Bee Paralysis Virus (ABPV), Chronic Bee Paralysis Virus (CBPV), in Vespa velutina specimens collected in Italy during 2017. Results of this investigation indicate that among pathogens, replicative form of KBV and BQCV were detected, assessing the spillover effect of both these viruses from managed honey bees to hornets.

Monitoring honey bee health in five natural protected areas in Italy.

The health status of the honey bee populations has attracted a great amount of interest in recent years. We investigated honey bee health in five natural protected areas in Italy from October 2009 to December 2010. Areas were selected to represent a wide range of bio­geographical zones including alpine, continental, and Mediterranean. Within each of these natural protected areas, one apiary of 20 colonies near potential pollution sources (e.g., agricultural areas, industrial areas, or urban settlements) and another apiary of 20 colonies far from possible sources of pollutants have been placed. To monitor honey bee health, colony mortality was related to: honey bee pathologies, environment (Naturality Index, plant protection products and heavy metal exposure), and apiary management. Anthropogenic pollutants and pathogens did not have significant effects on colony mortality while environment and the poor colony management skills of the beekeepers did.

Significance of the goby Zosterisessor ophiocephalus as a sentinel species for Venice Lagoon contamination: Combining biomarker responses and bioaccumulation.

The Venice Lagoon is an interesting example of an ecosystem suffering for a considerable anthropogenic impact, resulting in high concentrations of persistent organic pollutants (POPs) in lagoon sediments and seafood. In this context, biomonitoring is a crucially important task. The present study aimed at evaluating the validity of a multiple biomarker approach in a benthic fish species. A total of 567 Zosterisessor ophiocephalus (Gobiidae) fish were collected in spring and autumn from three areas of Venice Lagoon (Porto Marghera, Val di Brenta, and Cà Roman) showing high, intermediate and low amounts of POPs, respectively. Aryl hydrocarbon receptor (AHR) and cytochrome P450 1A (CYP1A) mRNA levels, CYP1A protein amount and ethoxyresorufin O-deethylase activity (EROD) were measured in pooled liver and gills (mRNA levels only). Such biological data were then compared with polychlorinated biphenyls (PCBs) residues, measured in grass goby muscle by gas chromatography. Aryl hydrocarbon receptor and CYP1A mRNAs, protein and EROD were upregulated in accordance with PCB amounts measured in Z. ophiocephalus muscles. In fact, the highest AHR and CYP1A induction was observed in fish sampled in close proximity of the industrial area of Porto Marghera. Overall, the present study confirm the grass goby as a reliable sentinel species for Venice Lagoon, and AHR/CYP1A/EROD as a sensitive set of biomarkers of exposure for AHR ligands.

Immunohistochemistry by anti-cleaved-Lamin A: an improved approach to tackle the misuse of glucocorticoids in cattle.

The illegal use of glucocorticoids (GCs) as growth-promoters (GPs) is prohibited in farm animals in the European Union because the strong pharmacological activity of most synthetic GCs produces residues that are dangerous for human consumption. Among the alternative methods proposed to increase the efficacy of official controls, histology was the technique of choice in Italy on account of its high performance level. The aim of this study was to evaluate the reliability of immunohistochemistry (IHC) using anti-cleaved-Lamin A antibody to enhance the performance of the histological test applied to GC-related microscopic changes in the thymus. Veal calves (VC) and beef cattle (BC) were raised and both underwent different growth-promoting protocols or were left untreated. The morphology of the thymus parenchyma was evaluated for cortical atrophy with concurrent adipose tissue infiltration, and a score of 1 to 3 was attributed. A semi-quantitative IHC analysis was also performed by counting the number of positive thymocytes in 5 randomly selected high-power fields (HPFs). The distribution of the thymus atrophy scores was significantly different among the subgroups in both BC and VC. The IHC values were higher in untreated than in treated animals, for both BC and VC. The association between thymus atrophy score and IHC positivity showed higher median values in control than in treated animals (independently of the treatment protocol), for both BC and VC. Our data shows that IHC against anti-cleaved-Lamin A antibody is a reliable marker to detect illegal GC treatments, administered either alone or in association with other growth promoters, in both BC and VC. Combining IHC with the thymus atrophy score improves the accuracy of the histological method in correctly identifying treated animals and could represent a valuable, reproducible method to be applied to large-scale screening programmes.