Search Results (51)

Male tract infections (MTIs) are a common clinical condition, often presenting without any signs nor symptoms of disease. As advised by the European Urology Guidelines dealing with this topic, patients are typically treated with antibiotics alone. Nevertheless, in between 40% and 50% of cases, antibiotic therapy is not effective in eradicating the semen infection. Therefore, persistent semen infection is frequently found upon semen culture evaluation following antibiotic therapy. In this study, we aimed to analyze the fecal microbiota of male infertile patients with persistent MTI in order to verify the prevalence of gut dysbiosis in these patients. We therefore enrolled 20 infertile patients with persistent MTIs after a proper cycle of antibiotic treatment. All patients performed the study for gut microbiota analysis after about 30 days after the last dose of antibiotic treatment. Gut microbiota analysis revealed that 50% of patients with persistent MTI presented a reduction in microbial biodiversity. Indeed, a situation of gut dysbiosis was reported in 75% of patients. In details, the Firmicutes–Bacteroidetes ratio was reduced in 70% of such patients, including 40% of patients where a severe reduction was observed due to an elevated abundance of Bacteroidetes (putrefactive dysbiosis). The most frequent enterotype was Prevotella-dominant (43%). We demonstrated for the first time that patients with recurrent MTIs have enterotypes associated with increased gut permeability and systemic inflammation. Further studies are required to analyze the molecular machinery by which gut dysbiosis exerts its role in patients with MTIs, in particular persistent MTIs, and how supplementation with probiotics might impact in terms of restoring eubiosis, in terms of eradicating the infection, and reducing prostate inflammation and eventually in terms of improving semen evaluation in male infertile patients. Full article

This study assessed and compared the postmortem concentrations of 6-monoacetylmorphine [6-MAM] and 6-acetylcodeine [6-AC], morphine, and codeine in various tissues and fluids from 52 postmortem cases related to heroin use. Samples were received at the Poison Control and Forensic Chemistry Center in Jeddah, Saudi Arabia, and analyzed using liquid chromatography–mass spectrometry. Descriptive and inferential statistics, including median, range, variability, and outliers, were used for analysis. The results showed significant variability in heroin and metabolite concentrations across different fluids and tissues. Tissue specimens were analyzed in 38 cases (73%), with 50% of cases exhibiting putrefaction. Blood and tissue samples were available in 39 cases, highlighting the need for alternative specimens in challenging cases. Notably, heroin metabolites were detected in unique matrices, such as nasal swabs, bladder tissues, lung tissues, and small intestine tissues, underscoring the potential of these samples in forensic investigations, especially when traditional bodily fluids are unavailable or compromised. These findings suggest that environmental factors, timing of substance use, and postmortem changes influence substance distribution, emphasizing the need to consider the location of death when interpreting toxicological results for accurate forensic analysis. This study provides valuable insights into the distribution, correlation, and significance of heroin and its metabolites in postmortem samples, aiding the confirmation of heroin overdose. These findings contribute to the limited data on postmortem cases in the Middle East and North Africa, particularly Saudi Arabia, supporting efforts to curb drug abuse in this region. This knowledge can inform public health strategies and forensic practices, ultimately aiding efforts to address and mitigate drug abuse. Full article

Cognitive enhancement through stimulants such as modafinil is becoming increasingly popular, with many individuals using prescription stimulants for non-medical purposes to improve alertness, attention, and mood. The misuse of such substances has raised concerns, particularly in forensic toxicology. The UHPLC-QqQ-MS/MS method was developed to quantify modafinil in evidentiary samples and biological materials. Additionally, the authors noted the presence of sodium adducts during the analysis of samples with high concentrations of modafinil. The method was validated for accuracy, precision, and linearity, with a concentration range of 0.1–10.0 µg/mL for the evidences and 1.0–100.0 ng/mL for blood. The method successfully detected modafinil as the sole substance in all evidences, with concentrations ranging from 90.7 to 120.8 mg, corresponding to 45.5% to 80.5% of the labeled dose. The method was applied to real post-mortem human cases, where, among others, the concentration of modafinil in blood was 110 ng/mL, whereas, in another case, the concentration of modafinil in the putrefaction fluid exceeded 1000 ng/mL. The developed UHPLC-QqQ-MS/MS method is effective for the quantification of modafinil in evidentiary samples and biological materials, offering a reliable tool for forensic toxicology applications. This method can be used to evaluate modafinil use in both legal and illicit contexts, including cases of overdose or misuse. Full article

This study aimed to investigate the effects of a Standardized Natural Citrus Extract (SNCE) on broiler chickens’ growth performance, gut health, carcass quality, and welfare. A total of 756 one-day-old Ross 308 males were randomly assigned to two groups: a control group (CTL) fed with a standard diet, and a citrus group (SNCE) fed with the same standard diet supplemented with 250 g/ton of feed of SNCE. Growth performance was recorded weekly until d 35, while mortality was recorded daily. The feed conversion ratio (FCR) and European Efficiency Index (EEI) of broiler chickens were also calculated weekly. At day 35, 10 birds per group were randomly selected for slaughter performance. In parallel, broiler chickens’ welfare was assessed according to the Welfare Quality Assessment Protocol. Caecal digest was also collected post mortem for short-chain fatty acids (SCFA) analyses, and jejunum samples were collected for ex vivo gut permeability assay. SNCE dietary supplementation enhanced broiler chickens’ performance, i.e., final bodyweight and EEI, compared to the CTL group. The carcass weight was also significantly higher in the SNCE group. In addition, the fat percentage was lower in the SNCE group. Regarding broiler chickens’ welfare and gut health parameters, Footpad Dermatitis (FPD) and gate score were also lower in birds supplemented with SNCE. The SCFA measurement showed a lower concentration of iso-butyric acid, iso-valeric acid, and total putrefactive SCFA in the SNCE group. The differences in gut permeability measured as TEER value indicate that using citrus extract lowered the risk of gut inflammation. This study provides valuable insights into the mechanisms of action that may underlie the observed effects of SNCE on performance, as demonstrated in this study and others. These effects could potentially be attributed to the reduction in inflammation and the enhanced utilization of nutrients. Further studies are needed to confirm these results. Full article

Plasma-treated water (PTW) recently entered science as a sanitizing agent, which possess the capability for on-demand production. It offers interesting possibilities for sustainable and resource-saving applications in healthcare and food production. The present study monitors the impact of PTW on suspended cells before the biofilm formation of the putrefactive bacterium B. subtilis. Light and electron microscope imaging captures the maturing of growing biofilms within the first 24 h. Microbiological assays (proliferation, LIVE/DEAD, and XTT), which mirror the proliferation of the bacterium, the metabolic activity, and the integrity of the cell membrane, underpinning the metabolic response of still-suspended cells. B. subtilis cells without any treatment build up a resistive biofilm within the 24 h. Cells that remain in the supernatant predominantly appear as monomers or dimers. Treated B. subtilis cells have hampered biofilm formation and were not able to build up a confluent growing biofilm within the first 24 h. Moreover, the microscopic observation of PTW-treated suspension showed cellular aggregates with an unusually high connectivity of the individual cells. The findings suggest this cellular reaction as a counter measure against the adverse impact of PTW treatment. The experiments show the adverse impact of PTW on B. subtilis–biofilm formation and the phenomenological reaction of B. subtilis. Full article

This study aimed to develop fecal low-odor adult dog feed using processed corn protein powder as the primary raw material and evaluate its effectiveness through feeding experiments. The objectives included analyzing the fundamental nutritional indicators, comparing the quality of fecal low-odor adult dog feed with commercially available dog feed, and assessing the changes in the eating behavior, physical condition, and physiological and biochemical indicators before and after feeding on the fecal low-odor adult dog feed. This study involved formulating dog feed using processed fermented corn protein powder and conducting nutritional analyses to compare moisture, crude protein, ash, crude fat, and starch contents. Physical properties such as hardness and cohesion were also evaluated. Feeding experiments were conducted with adult dogs to monitor the changes in the eating behavior and physical condition. Physiological and biochemical indicators, including ammoniacal nitrogen and putrefactive odor in feces, were measured both before and after feeding to assess the impact of the fecal low-odor adult dog feed. The analysis showed no significant difference in moisture (p > 0.05), crude protein (p > 0.05), ash (p > 0.05), crude fat (p > 0.05), and starch (p > 0.05) contents between the fecal low-odor adult dog feed and commercially available dog feed. Similarly, no significant difference was noted in the hardness (p > 0.05) and cohesion (p > 0.05) between the two types of dog feed. Both types of dog feed contained rich volatile compounds with varying compositions. The commercially available dog feed had a sour odor, whereas the fecal low-odor adult dog feed had a barbecue and resin-like smell. After feeding with the fecal low-odor adult dog feed, the liquid nitrogen content significantly decreased (p < 0.01). Also, the indole content, a major contributor to fecal odor, based on the data, decreased after feeding. Full article

In forensic investigation, determining the time and cause of death becomes challenging, especially in cases where the remains are found in advanced decomposition, rendering traditional toxicological samples unavailable or unreliable. Entomotoxicology, an emerging methodology within forensic science, leverages insect specimens collected from cadavers as alternative toxicological samples. Several laboratory and field research studies have highlighted the efficacy in detecting various drugs, toxins, and elements absorbed by insects feeding on cadaveric tissues, even at low concentrations. However, correlation studies between drug concentrations in conventional matrices and insects remain controversial due to unknown factors influencing drug metabolism and larval feeding activity. This paper presents four real cases in which human cadavers were discovered in advanced stages of decomposition, and toxicological analyses were performed on both insect samples and available matrices. The results presented complement the scant literature currently available on the application of entomotoxicology in real cases, providing insights into the correlation between larvae and human specimen results. Furthermore, guidelines to collect and preserve entomological evidence at the crime scene and during the autopsy for use in entomotoxicological analyses are provided. This advancement holds promise in aiding forensic investigations, particularly in cases where traditional methods cannot be applied or require supporting data for further validation. Full article

Once a silo has been opened, the silage inside will face challenges such as aerobic deterioration, rot, and contamination. Biocontrol bacteria, as a kind of biological antiseptic, are highly effective and natural and are gaining increasing attention. This study aimed to screen a strain with anti-microbial activity against silage spoilage microorganisms and examine its effects on the fermentation quality, aerobic stability, in vitro digestion, and methane production of silage. Lactic acid bacteria, pathogenic and rot-causing microorganisms, were used as indicators to screen the strains for putrefactive silage. The bacteriostatic spectrum, growth performance, and tolerance to the silage environment of the strain were tested. A strain named D-2 was screened from rotten whole-plant corn silage and identified as Bacillus velezensis through physiological and biochemical tests as well as 16S rDNA sequencing. This study found that D-2 exhibits antibacterial effects on several microorganisms, including Escherichia coli, Staphylococcus aureus, Salmonella enteritidis, Aspergillus niger, Saccharomyces cerevisiae, Fusarium oxysporum, and Fusarium graminearum. However, it has no adverse effect on Lactobacillus reuteri, Enterococcus faecium, or Lactobacillus casei. D-2 can attain a stable stage within 10 h and withstand temperatures of up to 70 °C. Moreover, this study found that D-2 had a high survival rate of over 97% after 48 h in a lactic acid environment with pH 4. Freshly chopped whole-plant corn was inoculated without or with D-2 and ensiled for 60 days. The results show that D-2 inoculations increase the content of water-soluble carbohydrates, acetic acid, and propionic acid in the silage and decrease the number of yeasts and molds, the NH₄⁺-N/TN ratio, and the pH. We also found that fermenting whole-plant corn with D-2 significantly increased the in vitro digestibility and the propionic acid content, while also significantly inhibiting methane production. After being exposed to air for 10 days, D-2 can still effectively reduce the total number of yeasts and molds, prevent the decrease in lactic acid bacteria, and inhibit the increase in the pH and NH₄⁺-N/TN ratio of silage products. Overall, D-2 is resistant to pathogenic and rot-causing microorganisms, allowing for easy adaptation to silage production conditions. D-2 can effectively improve aerobic stability and reduce losses in the nutritional value of silage, indicating possible applications for the prevention of silage rot and methane production. Full article

The consumption of “ready-to-cook” foods has been experiencing rapid expansion due to modern lifestyles, and they are often sold in economical multipacks. These foods necessitate packaging that maintains their quality for extended periods of time during home storage once the original packaging is opened. This study evaluates a chitosan-based film derived from low- and high-molecular-weight (MW) chitosan in acetic acid without synthetic additives as an alternative packaging material for “ready-to-cook” beef burgers. The burgers were stored at 8 °C after being removed from their sales packaging. A commercial polyethylene (PE) film designed for food use, devoid of polyvinylchloride (PVC) and additives, served as the reference material. The production of six biogenic amines (BAs), indicative of putrefactive processes, was monitored. Additionally, the release of four phthalates (PAEs), unintentionally present in the packaging films, was assessed using solid-phase microextraction coupled with gas chromatography/mass spectrometry (SPME-GC/MS). Microbiological tests were conducted to investigate the antimicrobial efficacy of the packaging against Aspergillus Niger NRR3112. The results showed that the chitosan-based films, particularly those with low MW (LMW), exhibited superior meat preservation compared to the PE films. Furthermore, they released PAEs below legal limits and demonstrated the complete inhibition of fungal growth. These findings highlight the potential of chitosan-based packaging as a viable and effective option for extending the shelf-life and maintaining the quality of “ready-to-cook” meat products during domestic storage. Full article

Bile has emerged as an alternative matrix for toxicological investigation of drugs in suspected forensic cases of overdose in adults and intoxications in children. Toxicological investigation consists in screening and, subsequently, confirming the result with specific techniques, such as liquid chromatography with tandem mass spectrometry (LC-MS/MS). As there is no screening test on the market to test postmortem bile specimens, the novelty of this study was in investigating the applicability of a chemiluminescence immunoassay, designed for other matrices and available on the market, on bile and validate its use, testing the agreement with LC-MS/MS analysis. Bile specimens were obtained from 25 forensic cases of suspected death from overdose and intoxication. Sample preparation for bile screening consists simply in centrifugation and dilution. Confirmation analysis allows simultaneous identification of 108 drugs and was validated on bile. Kappa analysis assessed a perfect agreement (0.81–1) between the assays for benzodiazepines, methadone, opiates, cocaine, oxycodone, cannabinoids, buprenorphine and pregabalin; a substantial agreement (0.41–0.6) was reported for barbiturates. No agreement was assessed for amphetamines, due to an abundance of putrefactive amines in postmortem specimens. In conclusion, this fast and easy immunoassay could be used for initial screening of bile specimens, identifying presence of drugs, except amphetamines, with reliability. Full article

Ethanol can be produced by many microorganisms that colonize a dead body. Ethanol’s concentration depends on the congener higher alcohols, 1-propanol, isobutanol, 2-methyl-1-butanol, 3-methyl-1-butanol, and 1-butanol, as previous research has shown. This correlation is expressed by mathematical models which estimate the concentration of microbial ethanol. The aim of this contribution was to study the ethanol and higher alcohols’ production in various laboratory bacterial and fungal cultures and the applicability of the bacterial and fungal models (which concern the bacteria E. coli, S. aureus, K. pneumoniae, and E. faecalis, and the fungus C. albicans) in these samples, as well as in blood samples from autopsy cases, with the overall objective of investigating the models’ applicability in routine casework. The bacteria and fungus were cultured in conventional culture media and in denatured human blood cultures under various conditions. The alcohols’ concentrations were determined using a head space–gas chromatography–flame ionization detector (HS-GC-FID). The previously reported bacterial and yeast models were applied in the cultured samples and in blood from 122 autopsy cases. Our results showed that 1-propanol was not produced by C. albicans and E. faecalis under certain conditions. Also, 1-butanol was not produced by C. albicans, E. faecalis, and K. pneumonia under certain conditions. Furthermore, the bacterial models were applicable in postmortem samples irrespective of the microbes that were possibly activated in the sample, while the EC models showed the best applicability among all the bacterial and yeast models. The best applicability of the bacterial models was observed in autopsy blood with 0.10 g/L < BAC < 1.0 g/L in cases of violent and undetermined causes of death and in cases with putrefaction. Finally, the yeast models were applicable in limited, possibly special, autopsy cases. In conclusion, it could be inferred that the source of ethanol in any given postmortem blood sample is likely microbial if either most bacterial models or at least one model from each distinct bacterial species is successfully applicable. Full article

The management of drainage water (DW), which is produced during the soilless cultivation of plants, requires a high energy input. At the same time, DW is characterized by a high electrolytic conductivity, a high redox potential, and is also stable and putrefaction-free. In the present study, the natural properties of drainage water and a biotreatment method employing an external organic substrate in the form of citric acid (C/N 1.0, 1.5, 2.0) were utilized for energy recovery by a microbial fuel cell (MFC). The cathode chamber served as a retention tank for DW with a carbon felt electrode fixed inside. In turn, a biological reactor with biomass attached to the filling in the form of carbon felt served as the anode chamber. The filling also played the role of an electrode. The chambers were combined by an ion exchange membrane, forming an H letter-shaped system. They were then connected in an external electrical circuit with a resistance of 1k Ω. The use of a flow-through system eliminated steps involving aeration and mixing of the chambers’ contents. Citric acid was found to be an efficient organic substrate. The voltage of the electric current increased from 44.34 ± 60.92 mV to 566.06 ± 2.47 mV for the organic substrate dose expressed by the C/N ratio ranging from 1.0 to 2.0. At the same time, the denitrification efficiency ranged from 51.47 ± 9.84 to 95.60 ± 1.99% and that of dephosphatation from 88.97 ± 2.41 to 90.48 ± 1.99% at C/N from 1.0 to 2.0. The conducted studies confirmed the possibility of recovering energy during the biological purification of drainage water in a biofilm reactor. The adopted solution only required the connection of electrodes and tanks with an ion-selective membrane. Further research should aim to biologically treat DW followed by identification of the feasibility of energy recovery by means of MFC. Full article

The development of food packaging materials that reduce the production of plastic, preserving at the same time the quality of food, is a topic of great interest today for the scientific community. Therefore, this article aims to report the effectiveness of an eco-friendly packaging material based on alginic acid and grape pomace extract from Vitis vinifera L. (winemaking by-products) for storing red meat in a domestic refrigerator. Specifically, biogenic amines are considered “sentinels” of the putrefactive processes, and their presence was thus monitored. For this purpose, an experimental analytical protocol based on the use of solid-phase microextraction coupled with gas chromatography–mass spectrometry was developed during this work for the determination of six biogenic amines (butylamine, cadaverine, isobutylamine, isopentylamine, putrescine, and tyramine). Moreover, by combining the analytical results with those of pH and weight loss measurements, differential scanning calorimetry, and microbiological analysis, it was proved that the studied materials could be proposed as an alternative packaging material for storing foods of animal origin, thus lowering the environmental impact according to sustainability principles. Full article

Performing quantitative sampling and determining faunistic analyses of dipterans is of fundamental importance in the analysis of ecological behavior, such as population dynamics and diversity, among other factors, for exotic and native species of necrophagous dipterans, so it is important to observe the type of bait used in traps to capture these dipteran species. This work aims to study structural parameters and faunistic indices of the diversity of Calliphoridae and Mesembrinellidae species as well as the abundance and diversity of species attracted to liver in two stages of decomposition: fresh liver and liver at 48 h of putrefaction. A total of 2826 dipterans were collected during the period from May 2021 to February 2022. We observed that liver in decomposition for 48 h was more attractive in the forest and rural environments, while fresh liver showed greater attractiveness in the urban environment; however, no statistical difference was evidenced between the attractiveness in the different environments. The Mesembrinellidae family and the species Lucilia eximia (Wiedemann, 1819) were collected mostly from deteriorated liver, while Cochliomyia macellaria (Fabricius, 1775) showed no preference for any liver decomposition stage. The Wilcoxon test indicated that there is a significant difference between the preferences for putrefied bait in Mesembrinellidae, while in Calliphoridae, there was no preference for type of bait. The faunistic analysis showed that richness in the forest area was always higher when compared to the urban and rural areas. Laneela nigripes (Guimarães, 1977) and Mesembrinella bellardiana (Aldrich, 1922) were abundant and exclusive in the preserved environment, showing themselves to be good environmental bioindicators. Full article

The pink teeth phenomenon has occasionally been observed in forensic autopsies. This study aimed to establish an experimental pink tooth model and an objective color tone evaluation method in order to clarify changes in the color tone of teeth and the relationship with hemoglobin monoxide and its decomposition products and with red pigment-producing bacteria, under various external environmental factors. It was confirmed that the color tone evaluation with ΔE and the L*C*h color space was useful. The results of various examinations using this model showed that color development was suppressed under aerobic conditions, faded early under light, became bright red under a low temperature and showed a tendency to be reddish at 3 days under high humidity and in the presence of soft tissue. The biochemical analysis revealed a significant increase in carboxyhemoglobin at 7 days and a tendency toward increasing the total heme pigment and bilirubin levels over time. The bacteriological analysis revealed that red pigment-producing bacteria increased over time but that the color faded after 7 days. These results suggest that putrefaction greatly affects the pink teeth phenomenon, whereas red pigment-producing bacteria have little effect on the occurrence of pink teeth. However, further studies are needed to clarify bacteriological involvement. Full article