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.

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.

The impact of full-fat Hermetia illucens larvae meal on the health and immune system function of broiler chickens.

Introduction: Alternative protein sources have recently been attracting growing interest as potential components of livestock nutrition. This study evaluated the effect on broiler health of replacing the soybean protein component of poultry feed with processed insect protein from farmed Hermetia illucens (HI) larvae. Material and Methods: A total of 384 male broiler chicks were divided into four diet groups (eight pens/treatment and 12 birds/pen) and reared to the 42nd day of life (dol). Each treatment group received a starter diet until the 14th dol, then a grower diet until the 35th and finally a finisher diet until the 42nd. The soybean meal in standard diets was replaced with full-fat HI larvae meal in the following amounts: 0% for the control group HI-0, 50% for group HI-50, 75% for group HI-75, and 100% for group HI-100. At 1 dol, chicks were vaccinated against Marek's disease, coccidiosis, Newcastle disease and infectious bronchitis and at 7 dol against avian metapneumovirus infections using live-attenuated vaccines. Blood and spleen samples were collected at three and six weeks of age and analysed using ELISA, flow cytometry, haematology and biochemistry. Results: On the 42nd dol, as the content of larvae meal in the chickens' diets increased, the birds' body weights decreased significantly. The substitution of the protein source had no effect on the haematological markers. In chicks that received larvae meal, there was a decrease in creatine kinase activity and phosphorus levels and an increase in calcium and uric acid levels in serum. Raising the proportion of full-fat HI larvae meal in the diet raised the percentage of T CD3+CD8a+ cells and lowered that of T CD3+CD4+ cells in both sample types. Chickens fed larvae meal had significantly lower post-vaccination anti-infectious bronchitis virus antibody titres. Conclusion: The poorer production results and impaired health in experimental birds may indicate lower than 50% protein substitution with full-fat HI larvae meal to be optimal.

The Use of Chitin from the Molts of Mealworm (Tenebrio molitor) for the Removal of Anionic and Cationic Dyes from Aqueous Solutions.

The possibility of using chitin from the molts of an insect-ealworm (Tenebrio molitor) to remove anionic (RB5, RY84) and cationic dyes (BV10, BR46) from aqueous solutions was investigated. The scope of the research included, among others: Characteristics of chitin from mealworms (FTIR, SEM, pHPZC), the effect of pH on sorption efficiency, sorption kinetics (pseudo-first, pseudo-second order, intramolecular diffusion models) and the determination of the maximum sorption capacity (Langmuir and Freundlich models). The sorption efficiency of anionic dyes on chitin from mealworm was the highest at pH 2-3, and for cationic dyes at pH 6. The equilibrium time of sorption of anionic dyes was 240-300 min and for cationic dyes it was 180-240 min. The experimental data on dye sorption kinetics was best described by the pseudo-second order model. The maximum sorption capacity of chitin from the mealworm for the anionic dyes RB5 and RY84 was 121.15 mg/g and 138.55 mg/g, respectively, and was higher than with some carbon-based materials (literature data). In the case of cationic dyes, the sorption capacity of the tested chitin was lower and reached 3.22 mg/g and 59.56 mg/g for BV10 and BR46, respectively.

Effect of Mineral-Microbial Deodorizing Preparation on the Value of Poultry Manure as Soil Amendment.

Poultry farming involves the production of poultry manures (PMs), which, if properly managed, are excellent organic soil amendments. Poultry farms generally do not have adequate arable land, and therefore, valuable fertilizer becomes a problematic waste. During the production and storage of PMs, odorous VOCs, NH4, H2S, and potent greenhouse gases such as CH4, CO2 are emitted. It influences the productivity of poultry and negatively affects the working conditions of working staff. In the present study, mineral-microbial deodorizing preparations (MMDP) based on perlite and bentonite as well as the following microorganism strains Lactobacillus plantarum, Leuconostoc mesenteroides, Bacillus megaterium, B. subtilis, and Pseudomonas fluorescens were added to the litter of turkey broilers (TB) and egg-laying hens (LH). PMs were compared with treatments without the addition of MMDP, and maize, sunflower, and rapeseed forage crops were tested. The influence on soil parameters such as pH, EC, HAC, SBC, CEC, BS, Ntot, Ctot, and plant yield and parameter of photosynthesis, i.e., SPAD index, was tested. Soil amending with manure resulted in an increase in pH and a decrease in HAC; in addition, an increase in EC, which was counteracted by the addition of MMDP, was noted. MMDP positively affected parameters such as SBC, CEC, and BS. It was shown that PMs, with the addition of MMDP, improved crops' yield in the first year of the study, whereas this effect was not seen for the after-crop plants (lupine). The main 'added value' related to the usage of MMDP in poultry production is the improvement in the properties of PMs, which mainly had a positive effect on soil indicators.

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.

Occurrence and Genetic Correlations of Yersinia spp. Isolated from Commensal Rodents in Northeastern Poland.

Rodents can be a potential Yersinia spp. vector responsible for farm facilities contamination. The aim of the study was to determine the prevalence of Yersinia spp. in commensal rodents found in the farms and fodder factory areas to characterize the obtained isolates and epidemiological risk. Intestinal samples were subjected to bacteriological, bioserotype, and PCR examination for virulence markers ail, ystA, ystB, and inv presence. Yersinia spp. was isolated from 43 out of 244 (17.6%) rodents (Apodemus agrarius n = 132, Mus musculus n = 102, Apodemus sylvaticus n = 8, Rattus norvegicus n = 2). Y. enterocolitica was isolated from 41 rodents (16.8%), and from one Y. pseudotuberculosis and one Y. kristensenii. In three cases, two Y. enterocolitica isolates were obtained from one rodent. All Y. enetrocolitica contained ystB and belonged to biotype 1A, considered as potentially pathogenic. One isolate additionally had the ail gene typical for pathogenic strains. The sequence analysis of the ystB, ail, and inv fragments showed a high similarity to those from clinical cases. The current study revealed a high prevalence of Y. enetrocolitica among commensal rodents, but the classification of all of Y. enterocolitica isolates into biotype 1A and the sporadic isolation of Y. pseudotuberculosis do not indicate a high epidemiological risk.

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.

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.

Partial and Total Replacement of Soybean Meal with Full-Fat Black Soldier Fly (Hermetia illucens L.) Larvae Meal in Broiler Chicken Diets: Impact on Growth Performance, Carcass Quality and Meat Quality.

The aim of this study was to compare selected growth performance parameters and slaughter characteristics in broiler chickens fed diets with a different content of full-fat Hermetia illucens L. (HI) larvae meal. The experiment was performed on 384 male broiler chickens (Ross 308) reared to 42 d of age and assigned to four dietary treatments (HI0-control diet and diets where soybean meal protein (SBM) was replaced with HI protein in 50%, 75% and 100%, respectively). The final body weights of chickens were as follows: 3010.0 g (HI0), 2650.0 g (HI50), 2590.0 g (HI75) and 2375.0 g (H100, p < 0.05). The carcasses of chickens from the experimental groups contained less meat and more abdominal fat. The feed conversion ratio for the entire experimental period was similar in groups HI0, HI50 and HI75 and more desirable than in group HI100 (p < 0.05). The meat of broiler chickens from groups HI75 and HI100 was characterized by significantly (p < 0.05) lower juiciness and taste intensity than the meat of birds from groups HI0 and HI50. The replacement of SBM protein with full-fat HI larvae meal in broiler diets exceeding 50% significantly compromised the growth performance of birds and the carcass and meat quality.

Effects of a Low Dose of T-2 Toxin on the Percentage of T and B Lymphocytes and Cytokine Secretion in the Porcine Ileal Wall.

Plant materials used in the production of pig feed are frequently contaminated with mycotoxins. T-2 toxin is a secondary metabolite of selected Fusarium species, and it can exert a harmful influence on living organisms. Most mycotoxins enter the body via the gastrointestinal tract, and they can modulate the gut-associated lymphoid tissue (GALT) function. However, little is known about the influence of low T-2 toxin doses on GALT. Therefore, the aim of this study was to evaluate the effect of T-2 toxin administered at 50% of the lowest-observed-adverse-effect level (LOAEL) on the percentage of CD2+ T cells, CD4+ T helper cells, CD8+ cytotoxic T cells, CD4+CD8+ double-positive T cells, TCRγδ+ cells, CD5+CD8- B1 cells, and CD21+ B2 cells, and the secretion of proinflammatory (IFN-γ, IL-1ß, IL-2, IL-12/23p40, IL-17A), anti-inflammatory, and regulatory (IL-4, IL-10, TGF-ß) cytokines in the porcine ileal wall. The results of the study revealed that T-2 toxin disrupts the development of tolerance to food antigens by enhancing the secretion of proinflammatory and regulatory cytokines and decreasing the production of anti-inflammatory TGF-ß. T-2 toxin triggered the cellular response, which was manifested by an increase in the percentage of CD8+ T cells and a decrease in the percentage of B2 and Tγδ lymphocytes.

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.

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.

Cytotoxicity of Odorous Compounds from Poultry Manure.

Long-term exposure and inhalation of odorous compounds from poultry manure can be harmful to farm workers and the surrounding residents as well as animals. The aim of the present study was to determine the cytotoxicity and IC50 values of common odorous compounds such as ammonium, dimethylamine, trimethylamine, butyric acid, phenol, and indole in the chick liver hepatocellular carcinoma cell line LMH (Leghorn Male Hepatoma), in vitro, using MTT (3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide) and PrestoBlue cytotoxicity assays. The cells were microscopically examined for any morphological changes post treatment. Dimethylamine exhibited the strongest cytotoxic effect on LMH cells with an IC50 value of 0.06% and 0.04% after an exposure of 24 h and 48 h, respectively. Both ammonium and trimethylamine had comparable cytotoxicity and their IC50 values were 0.08% and 0.04% after 24 h and 48 h, respectively. Of note, indole had the lowest cytotoxicity as the majority of cells were viable even after 72 h exposure. Thus, the IC50 for indole was not calculated. Results achieved from both MTT and PrestoBlue assays were comparable. Moreover, the morphological changes induced by the tested odours in LMH cells resulted in monolayer destruction, cytoplasm vacuolisation, chromatin condensation, and changes in nucleus and cell shape. Our study showed harmful effects of odorous compounds in chick tissues.

Impact of a microbial-mineral biopreparation on microbial community and deodorization of manures.

The aim of this study was to determine the number of bacteria in poultry, cattle and swine manure in order to perform hygienization and deodorization using a microbial-mineral biopreparation. The highest number of bacteria was recorded in laying hens manure (5.1×10(10) cfu/g). It was noted that bacteria: coliforms, E. coli, Clostridium, Enterococcus number was reduced (1-2 log) after the biopreparation application. The investigated odorous compound concentrations were reduced with 34-78% efficiency, depending on the type of manure and odorant. All odorous compounds were efficiently reduced only in the case of laying hen manure.