Innovative Applications of Tenebrio molitor Larvae in the Production of Sustainable Meat Sausages: Quality and Safety Aspects.

With the world's population continuing to grow, ensuring sustainable protein sources for everyone is becoming increasingly challenging. Despite meat being considered unsustainable, people find it challenging to abstain from consuming it. However, one solution to this dilemma could be the incorporation of mealworms into conventional meat products, i.e., sausages. The incorporation of mealworms into sausage formulations appears to shift the fatty acid profile towards higher levels of monounsaturated fats and polyunsaturated fatty acids (PUFAs), particularly omega-3s, potentially enhancing the nutritional value and offering health benefits. Therefore, our study aimed to improve the nutritional value and safety parameters of traditional sausages by enriching them with the flour of mealworm larvae. For this purpose, the larvae were reared on a sustainable substrate with brewery by-products, brewer's yeast, and carrots. They were used frozen and freeze-dried in sausage recipes, replacing pork in different proportions. The analysis of the product's chemical safety parameters (biogenic amines, nitrates and nitrites, volatile fatty acids (FA), and peroxide) and nutritional value (including collagen, cholesterol, amino acids, FA, and hydroxyproline) was carried out in an accredited laboratory. The results of our study have demonstrated that the incorporation of mealworms into sausages, particularly through freeze-drying, increased fat content and enhanced the profile of FA, including omega-3s while reducing protein and cholesterol levels, and altering collagen content, suggesting improved nutritional value and potential health benefits without compromising the safety of the product. Therefore, we are highlighting that the addition of mealworms influences the quality of amino acids positively and maintains biogenic amine levels within safe limits, alongside a negligible impact on nitrates and nitrites and a reduction in peroxide values. These findings indicate an overall improvement in sausage quality and safety without compromising safety.

Comparative Evaluation of Diagnostic Methods for Subclinical Benign Prostatic Hyperplasia in Intact Breeding Male Dogs.

Benign prostatic hyperplasia (BPH) is an androgen-related non-neoplastic enlargement of the prostate gland that commonly affects both reproductive capabilities and the general health of intact dogs. The subclinical form of BPH can be challenging to diagnose due to a lack of clinical signs, even if rectal palpation is performed. Left untreated, this condition poses risks to the dogs' health and breeding status. This study, involving 65 male dogs, aimed to investigate subclinical BPH through rectal palpation, ultrasonography, and analysis of canine prostatic-specific esterase (CPSE). Of the participants, 35 had subclinical BPH, and 30 served as a healthy control group. Dogs suspected of subclinical BPH, as determined by examination results from ultrasonography and CPSE analysis, underwent fine needle aspiration (FNA) guided by ultrasound to enhance diagnostic precision. Findings revealed distinct differences in rectal palpation and ultrasonography between subclinical BPH and healthy dogs. This study established diagnostic thresholds based on prostatic volume and CPSE values and proposed new thresholds for subclinical BPH. Additionally, results showed that prostate gland volume depended on the weight and the age of the dog. In conclusion, early detection of this condition is possible through various examinations, such as changes in ultrasound features, CPSE levels, and rectal palpation. All together, these methods can aid practitioners in early detection of BPH and assist with scheduling screening programs for dogs, ultimately promoting their overall health and reproductive well-being. In conclusion, we advocate for routine, non-invasive prostate screenings in breeding males, underlining the effectiveness of a combination of various multiple techniques for early subclinical BPH detection.

Analysis of Morphological and Morphometric Changes in a Parenchymal Tissue after the Radiofrequency Ablation Procedure.

Background and Objectives: Prostate cancer is on the rise in the European Union, and radiofrequency ablation (RFA) is one of the minimally invasive treatment options used for its treatment. Therefore, the aim of this study was to investigate and analyze the effects of RFA on prostate tissues. Materials and Methods: A standard prostate RFA procedure was performed on 13 non-purebred dogs in three sessions: no cooling (NC), cooling with a 0.1% NaCl solution (C.01), and cooling using a 0.9% NaCl solution (C.09). Microtome-cut 2-3 µm sections of prostate samples were stained with hematoxylin and eosin and further examined. Results: A histopathologic evaluation identified four zones of exposure: direct, application, necrosis, and transitional, as the damage on tissues decreased going further from the ablation site. The areas and perimeters of these zones were calculated, and geometric shapes of ablative lesions were evaluated using the quotient formula. Areas and perimeters of prostate tissue lesions in the NC and C.09 sessions were of similar size, whereas those found in C.01 were statistically significantly smaller. Lesions observed in session C.01 were of the most regular geometric shape, while the most irregular ones were found in session C.09. The shapes of lesions closest to the ablation electrode were the most irregular, becoming more regular the further away from the electrode they were. Conclusions: Prostate RFA leads to tissue damage with distinct morphological zones. Notably, the prostate lesions were the smallest and the most regular in shape after RFA procedures using the 0.1% NaCl cooling solution. It can be argued that smaller ablation sites may result in smaller scars, thus allowing for faster tissue healing if the blood flow and innervation at the ablation site are not compromised.

Zearalenone and Its Metabolites in Blood Serum, Urine, and Milk of Dairy Cows.

After oral contamination, zearalenone (ZEN) is rapidly absorbed in organisms and can be detected in biological fluids. In this study, we investigated the metabolites of ZEN in the biological fluids of cows (blood, urine, milk). The study was divided into three stages: preparation (the first stage), investigation (the second stage), and final stage (the third stage). Samples of biological fluids were taken 7, 21, and 35 days after the beginning of the study. At the first stage and at the second stage, the cows were fed a total mixed ration (TMR) with naturally contaminated mycotoxin-zearalenone (500 ± 75 µg/kg). In the third stage, the cows were fed a TMR without mycotoxins. This study established that at the second stage, the alpha-zearalenol concentrations in the serum increased by 92% and the beta-zearalanol in the urine decreased by 48% compared to the first stage (p < 0.05). The beta-zearalenol and zearalanone concentrations in the urine were higher compared to that of the alpha-zearalenol. The zearalenone concentration in the milk at the second stage was 35% higher than at the first stage (p < 0.05). A significant negative correlation (r = −0.540) was determined between the beta-zearalenol and beta-zearalanol concentrations in the urine and the positive significant correlation (r = 0.826) between the beta-zearalanol and alpha-zearalenol concentrations in the serum (p < 0.05). During the study, it was determined that feeding cows for two weeks with a TMR without mycotoxins can reduce concentrations of alpha-zearalanol, beta-zearalenol, and beta-zearalanol in the biological fluids and can reduce the concentrations of ZEN in the milk, but does not reduce the concentration of zearalanone.

Morphometric analysis of the red fox molar teeth in Lithuania.

The objective of the study was to investigate the lengths and widths of maxillary (P4 , M1 and M2 ) and mandibular (M1 , M2 and M3 ) molar teeth, lengths of dental rows and dental features of sexual dimorphism in the red fox. The correlation analysis between teeth measurements and dental row length was included too. The osteometric analysis was performed by the A. von den Driesch (1976) method. A total of 230 skulls and mandibles were investigated (n = 230): 113 males (n = 113) and 117 females (n = 117). The findings of the research revealed that most dimensions are higher in males compared with females (p < 0.001). Contrarily, Wilks' Lambda analysis showed results in the interval from 0.821 to 1, which leads to the conclusion that a teeth measurement study is unreliable to determine the sex of the red fox. The trustworthy measurements were the length of the whole dental arcade in the maxilla and mandible (UDR and LDR); Wilks' Lambda UDR = 0.860; LDR = 0.821; p < 0.001). Evaluation of the correlation between measurements revealed that results vary from very weak (0.047) to moderate strength (0.543). There were no strongly correlating measurements found. The correlation between the upper and lower molars with the length of the teeth arcade is quite weak, and only solitary measurements (M2 , L and B) had a weak correlation.

Prevalence of Mycotoxins and Endotoxins in Total Mixed Rations and Different Types of Ensiled Forages for Dairy Cows in Lithuania.

In this study, 119 samples of total mixed rations and different types of ensiled forage (maize and grass silage, and haylage) collected in 2019-2020 from dairy farms in Lithuania were analyzed to evaluate the quantitative occurrence of mycotoxins and endotoxins. Samples were analyzed using high-performance liquid chromatography (HPLC) with a fluorescent (FLD) and an ultraviolet detector (UV) of mycotoxins and a detection assay based on the ELISA technology for endotoxins. The study included toxins regulated within the European Union (aflatoxin B1 (AFB1), zearalenone (ZEA), deoxynivalenol (DON) and T-2 toxin) and nonregulated toxins (endotoxins). Mycotoxin analysis showed that 49.58% of the samples out of 119 were positive for AFB1, 52.11% for ZEA and DON, 55.47% for T-2 toxin and 84.04% for endotoxins. In the contaminated samples, the highest mean values of AFB1 and T-2 toxin were determined in the grass silage samples, while ZEA and DON-were determined in the maize silage samples. Maize silage samples had the highest ZEA and DON concentrations, exceeding the EU maximum permissible concentration limits. In the haylage samples, AFB1 mycotoxin exceeded the maximum concentration limits. The highest mean value of endotoxins was determined in the total mixed rations samples. This is the first study to provide information about the concentrations of mycotoxins and endotoxins in total mixed rations and different types of ensiled forages for dairy cows in Lithuania.

Microbial Diversity and Antimicrobial Resistance Profile in Microbiota From Soils of Conventional and Organic Farming Systems.

Soil is one of the biggest reservoirs of microbial diversity, yet the processes that define the community dynamics are not fully understood. Apart from soil management being vital for agricultural purposes, it is also considered a favorable environment for the evolution and development of antimicrobial resistance, which is due to its high complexity and ongoing competition between the microorganisms. Different approaches to agricultural production might have specific outcomes for soil microbial community composition and antibiotic resistance phenotype. Therefore in this study we aimed to compare the soil microbiota and its resistome in conventional and organic farming systems that are continually influenced by the different treatment (inorganic fertilizers and pesticides vs. organic manure and no chemical pest management). The comparison of the soil microbial communities revealed no major differences among the main phyla of bacteria between the two farming styles with similar soil structure and pH. Only small differences between the lower taxa could be observed indicating that the soil community is stable, with minor shifts in composition being able to handle the different styles of treatment and fertilization. It is still unclear what level of intensity can change microbial composition but current conventional farming in Central Europe demonstrates acceptable level of intensity for soil bacterial communities. When the resistome of the soils was assessed by screening the total soil DNA for clinically relevant and soil-derived antibiotic resistance genes, a low variety of resistance determinants was detected (resistance to ß-lactams, aminoglycosides, tetracycline, erythromycin, and rifampicin) with no clear preference for the soil farming type. The same soil samples were also used to isolate antibiotic resistant cultivable bacteria, which were predominated by highly resistant isolates of Pseudomonas, Stenotrophomonas, Sphingobacterium and Chryseobacterium genera. The resistance of these isolates was largely dependent on the efflux mechanisms, the soil Pseudomonas spp. relying mostly on RND, while Stenotrophomonas spp. and Chryseobacterium spp. on RND and ABC transporters.

Prevalence, species distribution and antimicrobial resistance patterns of methicillin-resistant staphylococci in Lithuanian pet animals.

BACKGROUND: The bacterial genus Staphylococcus consists of many species that causes infections in pet animals. Antimicrobial resistant staphylococci cause infections that are difficult to treat and they are important from the point of one health perspective. The aim of this study was to determine the prevalence of methicillin-resistant Staphylococcus (MRS) species, including methicillin-resistant S. aureus (MRSA) in diseased pet animals (Group A) and kennel dogs (Group B) in Lithuania and to characterize the isolates according to their antimicrobial resistance. RESULTS: Twenty-one MRS isolates were obtained from 395 clinical samples (5.3 %; CI 95 % 3.5-8.0) of Group A animals. Sixteen, four and one isolates were from dogs, cats and a pet rabbit, respectively. The mecA gene was present in 20 isolates, whereas one isolate was positive for the mecC gene. Twenty-one MRS isolates (20.0 %; CI 95 % 13.5-28.6) were obtained from the vagina of female dogs (n = 105) (Group B). All isolates carried the mecA gene. Twelve MRS species were isolated of which S. pseudintermedius was the most common (18/42) followed by S. haemolyticus (8/42) and S. lentus (4/42). MRSA was not found. All MRS strains were susceptible to vancomycin, linezolid, daptomycin and quinupristin/dalfopristin. Resistance to tetracycline (16/21), clindamycin (15/21) and erythromycin (14/21) was the most common types of resistance in Group A animals. Three isolates also demonstrated resistance to rifampin. Resistance toward gentamicin (16/21), ciprofloxacin (15/21), macrolides (15/21) and tetracycline (12/21) was the most common in kennel dogs (Group B). The most common genes encoding resistance to antimicrobials (excluding beta-lactams) in isolates from Group A pets were tetK (21/42), aph(3')-IIIa (11/42) and aac(6')-Ie-aph(2'')-Ia (9/42). CONCLUSIONS: A wide range of MRS species were found in pet animals in Lithuania. MRSA was not found.