Foodborne Diseases in the Edible Insect Industry in Europe-New Challenges and Old Problems.

Insects play a key role in European agroecosystems. Insects provide important ecosystem services and make a significant contribution to the food chain, sustainable agriculture, the farm-to-fork (F2F) strategy, and the European Green Deal. Edible insects are regarded as a sustainable alternative to livestock, but their microbiological safety for consumers has not yet been fully clarified. The aim of this article is to describe the role of edible insects in the F2F approach, to discuss the latest veterinary guidelines concerning consumption of insect-based foods, and to analyze the biological, chemical, and physical hazards associated with edible insect farming and processing. Five groups of biological risk factors, ten groups of chemical risk factors, and thirteen groups of physical risks factors have been identified and divided into sub-groups. The presented risk maps can facilitate identification of potential threats, such as foodborne pathogens in various insect species and insect-based foods. Ensuring safety of insect-based foods, including effective control of foodborne diseases, will be a significant milestone on the path to maintaining a sustainable food chain in line with the F2F strategy and EU policies. Edible insects constitute a new category of farmed animals and a novel link in the food chain, but their production poses the same problems and challenges that are encountered in conventional livestock rearing and meat production.

Edible Insect Farming in the Context of the EU Regulations and Marketing-An Overview.

Insects are increasingly being considered as an attractive source of protein that can cater to the growing demand for food around the world and promote the development of sustainable food systems. Commercial insect farms have been established in various countries, mainly in Asia, but in Europe, edible insects have not yet emerged as a viable alternative to traditional plant- and animal-based sources of protein. In this paper, we present an interdisciplinary overview of the technological aspects of edible insect farming in the context of the EU regulations and marketing. Based on a review of the literature, we have concluded that edible insect farming can be a viable business sector that significantly contributes to the overall sustainability of food systems if the appropriate regulations are introduced and food safety standards are guaranteed. However, the success of the edible insect industry also requires consumer acceptance of entomophagy, which is rather low in Western societies. Therefore, targeted marketing strategies are indispensable to support the implementation of edible insect programs.

Molecular Detection of Selected Pathogens with Zoonotic Potential in Deer Keds (Lipoptena fortisetosa).

Deer keds are obligatory hematophagous ectoparasites of birds and mammals. Cervids serve as specific hosts for these insects. However, ked infestations have been observed in non-specific hosts, including humans, companion animals, and livestock. Lipoptena fortisetosa is a weakly studied ectoparasite, but there is evidence to indicate that it continues to spread across Europe. The existing knowledge on deer keds' impact on wildlife is superficial, and their veterinary importance is enigmatic. Lipoptena fortisetosa is a species with vectorial capacity, but potential pathogen transmission has not been assessed. The objective of this study was to evaluate the prevalence of selected pathogens in L. fortisetosa collected from cervids and host-seeking individuals in the environment. Out of 500 acquired samples, 307 (61.4%) had genetic material from at least one tested pathogen. Our research suggests that L. fortisetosa may be a potential vector of several pathogens, including A. phagocytophilum, Babesia spp., Bartonella spp., Borellia spp., Coxiella-like endosymbionts, Francisiella tularensis, Mycoplasma spp., Rickettsia spp., and Theileria spp.; however, further, more extensive investigations are required to confirm this. The results of the study indicate that keds can be used as biological markers for investigating the prevalence of vector-borne diseases in the population of free-ranging cervids.

Molecular Characterization of Lipoptena fortisetosa from Environmental Samples Collected in North-Eastern Poland.

Recent years have witnessed an increase in the population of Lipoptenafortisetosa in Central Europe. The genetic profile of this ectoparasite has not been studied in Poland to date. The aim of the present study was to confirm the presence of L.fortisetosa in north-eastern Poland and to characterize the examined population with the use of molecular methods. Deer keds were collected between June and July 2019 in six natural, mixed forests. A fragment of the rRNA 16S gene was used as a marker to identify L.fortisetosa by polymerase chain reaction (PCR). DNA samples were sequenced in the last step. Six new locations of L. fortisetosa were confirmed. No significant differences were observed in the sex ratios of L. cervi and L. fortisetosa (L. cervi p-value = 0.74; L. fortisetosa p-value = 0.65). Significant differences were noted between the total size of L. cervi and L. fortisetosa populations (p-value < 0.001). The similarity to GenBank sequences ranged from 95.56% to 100%. The obtained nucleotide sequences were very closely related to L. fortisetosa sequences from Lithuania, the Czech Republic, and Japan. Molecular analyses revealed considerable genetic diversity, which could indicate that various ectoparasite lineages have spread throughout Europe.

The merits of entomophagy in the post COVID-19 world.

BACKGROUND: In 2020, human society underwent several drastic changes due to the coronavirus disease 2019 (COVID-19) pandemic, which generated an unprecedented global impact. Since the onset of the COVID-19 pandemic, various pressing concerns underlying food security, such as transport, production, and maintenance of the supply chain, have been raised. SCOPE AND APPROACH: The present study aimed to describe and review the merits of entomophagy in the post COVID-19 world, especially with regard to the low risk for zoonotic disease spread, high production rate, and future prospects for inducing entomophagy to enhance the diversity in the food system in comparison to conventional livestock. KEY FINDINGS AND CONCLUSIONS: The advantages of entomophagy in the post COVID-19 world have been elucidated herein, with particular emphasis on the minimal risk of zoonotic disease transmission and production efficiency, in addition to the future goal of establishment of entomophagy to expand redundancy and diversity in the food system as against the utility of conventional livestock. In the current scenario, as well as in the post COVID-19 situation, boosting entomophagy may play a pivotal role in global food security, as the aspects already touched upon have amply demonstrated.

Molecular characterization of Lipoptena cervi from environmental samples collected in Poland.

The activity of Lipoptena cervi has intensified in Poland in recent years. The population genetics of this ectoparasite in Poland has never been described in the literature. The objectives of this study were to investigate the population genetics of L. cervi in selected regions of Poland, to evaluate molecular differences between L. cervi populations, and to determine phylogenetic relationships with other L. cervi sequences obtained in previous studies. In 2019, louse flies were sampled in natural mixed forests in five Polish voivodeships. Seven samples of L. cervi were collected from each voivodeship, and a total of 35 insects were analyzed molecularly. In the first step, Lipoptena spp. were identified to species level under a stereoscopic microscope. A fragment of the rRNA 16S gene was used as a marker to identify L. cervi by the PCR assay. The sequences were assigned accession numbers MT337409 to MT337416. A total of eight haplotypes were identified, two of which were dominant. In the obtained sequences, intraspecific pairwise genetic distances varied between 0.000 and 0.0496 (m = 0.0135; SD = 0.0149; SE = 0.0006; V = 110.11). Mean interpopulation diversity was d = 0.0135 (SE = 0.0027). The acquired nucleotide sequences were highly similar to the sequences from the Czech Republic (MF495940, AF322437), Lithuania (MN889542-MN889544) and Poland (MF541726-MF541729). The similarity with GenBank sequences ranged from 97.24% to 100%. This study revealed two dominant haplotypes of L. cervi in Poland, MT337410 and MT337413. Fragments of the analyzed sequences were detected in only one voivodeship. These findings suggest that the two dominant sequences are the oldest sequences that gave rise to the locally identified haplotypes. The lack of significant correlations with the sequences obtained in regions situated west of the research sites suggests the presence of other genetic populations in Europe.

A Comparative Analysis of Selected Methods for Determining Young's Modulus in Polylactic Acid Samples Manufactured with the FDM Method.

The objective of this study was to compare three methods for determining the Young's modulus of polylactic acid (PLA) and acrylonitrile-butadiene-styrene (ABS) samples. The samples were manufactured viathe fused filament fabrication/fused deposition modeling (FFF/FDM) 3D printing technique. Samples for analysis were obtained at processing temperatures of 180 °C to 230 °C. Measurements were performed with the use of two nondestructive techniques: the impulse excitation technique (IET) and the ultrasonic (US) method. The results were compared with values obtained in static tensile tests (STT), which ranged from 2.06 ± 0.03 to 2.15 ± 0.05 GPa. Similar changes in Young's modulus were observed in response to the processing temperatures of the compared methods. The values generated by the US method were closer to the results of the STT, but still diverged considerably, and the error exceeded 10% in all cases. Based on the present findings, it might be concluded that the results of destructive and nondestructive tests differ by approximately 1 GPa.

Temporal-Microclimatic Factors Affect the Phenology of Lipoptena fortisetosa in Central European Forests.

The objective of this study was to determine the correlations between the abundance of Lipoptena fortisetosa on new potential hosts and selected temporal-microclimatic conditions in a forest at the beginning of the host-seeking period. Louse flies were collected between 6 May and 15 July of 2019 and 2020 in a natural mixed forest in Poland. Keds were collected by three investigators walking along the same forest route during each sampling session. The number of captured keds and the date (time), temperature (°C), relative humidity (%), air pressure (hPa) and wind speed (km/h) were recorded. A total of five measurements were performed during each sampling session. The influence of temporal-microclimatic conditions on the number of collected ectoparasites was evaluated with the use of a Generalized Additive Model (GAM). A total of 1995 individuals were obtained during field surveys. The results of the GAM revealed a correlation between the number of host seeking L. fortisetosa vs. time, temperature, relative humidity, and wind speed. An increase in temperature was most highly correlated with the abundance of louse flies in the environment.

Gastric canthariasis caused by invasion of mealworm beetle larvae in weaned pigs in large-scale farming.

BACKGROUND: Mealworm beetle T. molitor (Coleoptera: Tenebrionidae) (Linnaeus, 1758) is one of the most important cosmopolitan primary storage pests, scavenging on a variety of post-harvest grains and affecting the quality and safety of food and feed. In addition to being an important factor in feed hygiene, the insect can also be an epidemiological factor of canthariasis. Livestock infestations with T. molitor are rarely reported. This article describes T. molitor-caused canthariasis in pigs in large scale closed-cycle farming. RESULTS: In the spring, we registered a significantly increased mortality among weaned pigs. In autopsy, live 3-6 mm long T. molitor larvae were found in their stomachs, especially in the non-glandular oesophageal region, on average 2-3 larvae per 10 cm2 of gastric mucosa. Corrective actions reduced the number of deaths back to basal levels. CONCLUSIONS: This is the first documented case of potentially lethal gastric canthariasis in weaned pigs, caused by invasion of T. molitor larvae. Although canthariasis caused by T. molitor has not been a significant problem in farm animals so far, our case indicates that the presence of mealworm beetles is a potential threat to animal welfare and health.

Internal parasites infecting dogs in rural areas

Dogs which spend their lives in rural areas are primarily "sentinels" of animal herds. The dogs' close contact with humans and other animals obligates their caregivers to systematic deworming. The goal is to eliminate potential contamination with parasites which is mostly caused by direct contact or food and water contaminated with dogs' excretions. The aim of this study was to assess internal parasitic invasions on dogs from rural areas which spend most of their lives on farms. In total, 69 samples of fresh stools were collected for the analysis, including 26 from females and 43 from males. Coprological analysis of stool samples was accompanied with a survey where pet owners gave information on the number of visits to the veterinarian in suspicion of their dogs being infected with the parasites, the number of deworming treatments in the previous year and the dogs' behavior towards strangers. Also, the age, sex and body weight of dogs were recorded. Statistical analysis of the obtained results was performed using Statistica 12.5 with a medical set by Statsoft. In total, 29 (46,03%) out of 69 fecal samples contained developmental forms of parasites. Research has shown that animals most vulnerable to invasion were in the 6­10 years old age group, mostly males. In addition, the invasion of Toxocara canis roundworms occurred most frequently, as it was found in 12 (41.37%) animals (6 females and 6 males). The analysis of the questionnaires showed that the dog owners' knowledge of the risk of infection and parasites is unsatisfactory. This corresponded with a small number of deworming treatments. It is significant that nearly half of the owners never dewormed their dogs.

A parasitological evaluation of edible insects and their role in the transmission of parasitic diseases to humans and animals.

From 1 January 2018 came into force Regulation (EU) 2015/2238 of the European Parliament and of the Council of 25 November 2015, introducing the concept of "novel foods", including insects and their parts. One of the most commonly used species of insects are: mealworms (Tenebrio molitor), house crickets (Acheta domesticus), cockroaches (Blattodea) and migratory locusts (Locusta migrans). In this context, the unfathomable issue is the role of edible insects in transmitting parasitic diseases that can cause significant losses in their breeding and may pose a threat to humans and animals. The aim of this study was to identify and evaluate the developmental forms of parasites colonizing edible insects in household farms and pet stores in Central Europe and to determine the potential risk of parasitic infections for humans and animals. The experimental material comprised samples of live insects (imagines) from 300 household farms and pet stores, including 75 mealworm farms, 75 house cricket farms, 75 Madagascar hissing cockroach farms and 75 migrating locust farms. Parasites were detected in 244 (81.33%) out of 300 (100%) examined insect farms. In 206 (68.67%) of the cases, the identified parasites were pathogenic for insects only; in 106 (35.33%) cases, parasites were potentially parasitic for animals; and in 91 (30.33%) cases, parasites were potentially pathogenic for humans. Edible insects are an underestimated reservoir of human and animal parasites. Our research indicates the important role of these insects in the epidemiology of parasites pathogenic to vertebrates. Conducted parasitological examination suggests that edible insects may be the most important parasite vector for domestic insectivorous animals. According to our studies the future research should focus on the need for constant monitoring of studied insect farms for pathogens, thus increasing food and feed safety.

The resistance of Eimeria spp. to toltrazuril in black grouse (Lyrurus tetrix) kept in an aviary.

The black grouse is a critically endangered species in Europe. Aviary breeding and re-introduction programs have contributed to the reinstatement of black grouse populations. Parasitic diseases can influence the species' behavior, choice of habitats, and survival rates. Protozoa of the genus Eimeria are the most prevalent parasites in black grouse. The aim of this study was to evaluate the effectiveness of toltrazuril (TOL) against Eimeria spp. in black grouse kept in an aviary. Eimeria spp. oocysts (mean OPG for all birds = 7,047.84; SD = 5,625.05) were detected in feces samples in a routine parasitological examination. Parasitic infections were treated with toltrazuril Baycox 2.5% (Bayer, Leverkusen, Germany) at 1 ml/1 l H2O administered per os with drinking water twice a day for 2 d, 12 h/24 h. High oocysts counts persisted upon the second parasitic examination. Therefore, the birds were divided into 3 groups in 3 existing aviaries. Toltrazuril was administered according to the same protocol, and its uptake with drinking water was monitored. A parasitological examination conducted 5 d later revealed the presence of parasites, and the TOL dose was increased to 3 ml/1 l H2O. When oocysts were detected in a successive exam, the TOL dose was further increased to 5 ml/1 l H2O. The increase in TOL dose did not eliminate parasites, therefore, the treatment was discontinued. The fecal egg count reduction test (FECRT) revealed that 1 ml of TOL reduced fecal oocysts counts (OPG) by 0.02%, the second application of the same TOL dose reduced OPG by 0%, 3 ml of TOL reduced OPG by 0%, and 5 ml of TOL reduced OPG by 65.7%. The applied doses of TOL did not induce significant differences in the fecal oocysts counts of any of the identified species of coccidia. The results indicate that the coccidia species infecting black grouse have natural resistance to TOL.

Treatment of cryptosporidiosis in captive green iguanas (Iguana iguana).

There are no standard guidelines for the treatment of cryptosporidiosis in reptiles. The aim of this study was to evaluate the efficacy of two cryptosporidiosis therapies in captive green iguanas. Eight green iguanas aged 2-6 years, including 6 (1 ♂ and 5 ♀) animals with chronic diarrhea, received treatment for cryptosporidiosis. The presence of Cryptosporidium sp. oocysts was determined in 8 iguanas (100%), Isospora sp. oocysts were detected in 3 animals (37.5%), and Oxyuridae eggs were observed in 5 iguanas (62.5%). The animals were divided into two therapeutic groups (A and B). Group A iguanas were administered halofuginone (Halocur, 0,50 mg/ml Intervet Productions S.A., France) at a dose of 110 mg/kg body weight (BW) every 7 days for 5 weeks. Group B animals were administered sulfadiazine and trimethoprim (Norodine Vet Oral Paste sulfadiazine 288,3 mg/g, trimethoprim 58 mg/g, ScanVet Animal Health A/S, Denmark) at 75 mg/kg BW per os every 5 days for 5 weeks and spiramycin and metronidazole (Stomorgyl, spiramycin 1500000 IU, metronidazole 250 mg, Merial, France) at 200 mg/kg BW every 5 days for 5 weeks. Both groups received hyperimmune bovine colostrum and subcutaneous fluids. Before treatment, the average number of Cryptosporidium sp. oocysts in 1 g of feces was determined at 1.71 * 105 (±313,262.44) in group A and 1.56 * 105 (±262,908.53) in group B; the average number of Isospora sp. oocysts was determined at 3.53 * 103 (±1747.38), and the average number of Oxyuridae eggs was determined at 810 (±496.74). Blood tests were performed once before treatment. The results of blood morphology and biochemistry tests before treatment revealed leukocytosis with a significant increase in heterophile and monocyte counts in all animals. Dehydration, elevated hematocrit values and low levels of Na+, Ca2+, PO4- and Cl- ions were observed in 6 iguanas. Two iguanas died during treatment. The gross necropsy revealed acute inflammation of gastric and duodenal mucosa, mucosal ecchymoses in the gastrointestinal tract, hepatomegaly and liver congestion, cholecystitis, enlarged kidneys and renal edema and congestion, cystitis, and an absence of fat bodies. Parasites were not detected in any developmental form after 40 days of therapy and during an monthly 18-month follow-up period. Effective treatment of cryptosporidiosis in reptiles minimizes the adverse consequences of disease, improves the animals' well-being and decreases euthanasia rates.

Odorous Compounds from Poultry Manure Induce DNA Damage, Nuclear Changes, and Decrease Cell Membrane Integrity in Chicken Liver Hepatocellular Carcinoma Cells.

Animal breeding and management of organic wastes pose a serious problem to the health of livestock and workers, as well as the nearby residents. The aim of the present study was to determine the mechanisms of toxicity of selected common odorous compounds from poultry manure, including ammonia, dimethylamine (DMA), trimethylamine (TMA), butyric acid, phenol, and indole. We measured their genotoxic and cytotoxic activity in the model chicken cell line (LMH), in vitro, by comet assay and lactate dehydrogenase assay, respectively. We also made microscopic observations of any morphological changes in these cells by DAPI staining. Four compounds, namely ammonia, DMA, TMA, and butyric acid increased DNA damage in a dose-dependent manner (p < 0.05), reaching genotoxicity as high as 73.2 ± 1.9%. Phenol and indole induced extensive DNA damage independent of the concentration used. Ammonia, DMA, and TMA caused a dose-dependent release of lactate dehydrogenase (p < 0.05). The IC50 values were 0.02%, 0.05%, and 0.1% for DMA, ammonia and TMA, respectively. These compounds also induced nuclear morphological changes, such as chromatin condensation, shrinkage, nuclear fragmentation (apoptotic bodies), and chromatin lysis. Our study exhibited the damaging effects of odorous compounds in chick LMH cell line.

Tongue worm (Pentastomida) infection in ball pythons (Python regius) ­ a case report

Tongue worms (Pentastomida) are endoparasites causing pentastomiasis, an invasive disease representing a threat to exotic animals and humans. Animals acquire infection via the alimentary tract. In reptiles, the parasite is present in the lungs, resulting in symptoms from the respiratory system. Pentastomiasis may be asymptomatic, but nonspecific symptoms may occur at high parasite concentrations. Due to the harmful effects of many antiparasitic substances, tongue worm invasion in reptiles remains not fully treatable. Although pentasomiasis is rarely diagnosed in Poland, pentastomids were diagnosed in two ball pythons, who were patients of the "Poliklinika Weterynaryjna" veterinary clinic. They demonstrated problems with the respiratory system and a significant deterioration of health. Fenbendazole at a dose of 100 mg/kg b.w., repeated after 7 days was shown to be effective.

Parasites of wild animals as a potential source of hazard to humans.

The decline in wild animal habitats and the uncontrolled growth of their population make these animals come closer to human settlements. The aim of the study was to identify parasitic infections in wild animals in the selected area, and to specify the hazards they create for humans. In more than 66% of the analysed faecal samples from wild boar, hares, roe deer, deer and fallow deer various developmental forms of parasites were found. These included parasites dangerous for humans: Toxocara canis, Capillaria hepatica, Capillaria bovis, Trichuris suis, Trichuris ovis, Trichuris globulosus, Eimeria spp., and Trichostongylus spp. It is necessary to monitor parasitic diseases in wild animals as they can lead to the spread of parasites creating a hazard to humans, pets and livestock.