A novel kit for enrichment of fecal helminth eggs.

We developed a new concentration kit, called the ParaEgg (PE), for easy detection trematode eggs from fecal samples in endemic areas of clonorchiasis and metagonimiasis in Korea. To create a standard of detection efficiency, 120 fecal samples were examined using the water-ether concentration method (WECM). The PE kit and Mini ParaSep (PS) kit were used to compare the detection sensitivity of 100 egg-positive and 20 egg-negative samples in WECM. Additionally, stool samples, which were intentionally spiked with 10, 20, and 30 Clonorchis sinensis eggs, were evaluated to assess the sensitivity in lowinfection cases. The PE and PS kits showed detection rates of 100% and 92%, respectively, from 100 egg-positive samples in WECM. Meanwhile, eggs were detected in 3 (PE) and 2 (PS) out of 20 egg-negative samples in WECM. The PE kit detected the highest number of eggs per gram of feces (727 on average), followed by the WECM (524) and PS kit (432). In fecal samples that were intentionally spiked with 10, 20, and 30 C. sinensis eggs, PE only detected eggs 2 out of 5 samples in 10 eggs spiked (40%), and the detection rates were 80% and 100%, respectively. The PE kit enabled a more accurate identification of trematode eggs because of the clearance of small fecal debris in the microscopic field. In conclusion, the PE kit is obviously helpful to detect and identify trematode eggs in stool examinations especially in endemic areas of clonorchiasis and metagonimiasis.

In vitro production of viable eggs from undeveloped oocytes in mouse preantral follicles by reconstructing granulosa cell-oocyte complexes†.

In vitro culture of ungrown oocytes in preantral follicles is one of the intriguing subjects being pursued to produce viable eggs in assisted reproductive technology. Previous studies have succeeded in obtaining mature eggs after in vitro culture of preantral follicles, while denuded undeveloped oocytes, which are obtained occasionally when collecting preantral follicles, seem to be almost useless. Moreover, methods to culture them efficiently to produce viable eggs have not been established yet. The present study was conducted to demonstrate in vitro culture of mouse denuded undeveloped oocytes by reconstructing granulosa cell-oocyte complexes, and to analyze cellular communication in reconstructed granulosa cell-oocyte complexes. Single denuded undeveloped oocytes were aggregated with 1 × 104 granulosa cells in wells with U-shaped bottoms in a low-binding cell culture plate for 8 days under either 20% or 5% O2, and then the reconstructed granulosa cell-oocyte complexes formed were cultured on a collagen-coated culture membrane insert for 4 days under 5% O2. At day 8 of culture, the rates of reconstructed granulosa cell-oocyte complexes formation were significantly higher in the culture group under 5% O2 (64.9%) than that under 20% O2 (42.3%; P < 0.001); furthermore, the formation of transzonal projections was observed. After maturation and fertilization, we produced matured eggs and blastocysts at higher rates (>90% and 61.9%, respectively) in the group cultured under 5% O2. After transferring 126 two- to four-cell stage embryos, six live pups were obtained. This is the first report that demonstrates production of viable eggs after in vitro culture of denuded undeveloped oocytes from preantral follicles by reconstruction of granulosa cell-oocyte complexes.

Levels of PCDD/Fs, PCBs, metals and rare earth elements in eggs and vegetables from areas with different environmental contamination impacts in the Campania region (Southern Italy).

Studying the links between environmental pollution and the levels of contamination in food is an important challenge to ensure human health. Matched samples of eggs from free-range hens and vegetables were analysed to investigate the bioaccumulation of PCDD/Fs, PCBs, metals and rare earth elements. Only two egg samples resulted above the limit fixed for PCDD/Fs and the action level set for DL-PCBs. The highest concentrations were found in the eggs from an area situated in a big city affected by strong urbanisation. Although eggs and vegetables were subjected to the same environmental pollution, the PCDD/F and PCB bioaccumulation that occurred in the eggs was much higher than those in vegetables (p < 0.01). In vegetables, the highest PCDD/F and PCB concentrations were found in lettuce and potatoes grown on contaminated soil. Higher bioaccumulation of Fe and Zn occurred in eggs compared to vegetables; La, Pr, Nd, Sm and Eu were found only in lettuce samples. The results of this study may provide important data useful in the risk assessment of human exposure through diet in accidents involving dangerous chemicals. Furthermore, the estimated weekly intakes calculated for PCDD/Fs and PCBs highlighted that, although vegetables accumulate very low concentrations of these contaminants, they contribute more than eggs to human exposure.

Mating of unfed, engorged, and partially to fully gravid Aedes aegypti (Diptera: Culicidae) female mosquitoes in producing viable eggs.

BACKGROUND: Understanding the relationship between blood-feeding and mating is important in effectively managing the most well-adapted vector insect, Aedes aegypti (Linnaeus). Although extensive studies have investigated the behavioural aspects of Aedes such as blood-feeding, mating, and their relationship, several knowledge gaps still exist. Therefore, the present study was undertaken to determine the possibility of successful mating by unfed, engorged, and partially to fully gravid (up to 5 days after blood-feeding with fully developed eggs) female Ae. aegypti mosquitoes and production of viable eggs. METHODS: Mating of sexually mature adult Aedes aegypti was allowed in three different ways. In control 1, the females were allowed to mate before taking blood meal, and in control 2, the females were not at all allowed to mate. In the experiment, the females were separated into six categories, viz. D-0 to D-5. In D-0, the females were allowed to mate immediately after the bloodmeal and, in D-1, the females were allowed to mate on the first day of blood feeding, likewise, the females of D-2, D-3, D-4 and D-5 were allowed to mate on 2nd, 3rd, 4th and 5th day of blood feeding. Ovitrap was uniformly kept on the 4th day of blood feeding for the cages D-0 to D-3 for 1 h and then removed and for the cages D-4, and D-5, the ovitrap was kept on 4th and 5th day of blood feeding for 1h immediately after mating. The total number of eggs and the total number of hatching were counted. In the subsequent days, the entire experiment was replicated two times with different cohorts of mosquitoes, and the mean value of three experiments was used to draw Excel bars with 5% error bars and also for the statistical analysis. RESULTS: It was found that mating just before oviposition was sufficient to produce 1581 eggs (70% compared with control) and fertilize 1369 eggs (85% compared with total eggs laid), which is far higher than the 676 non-hatching (unfertilized) eggs (30%) laid by unmated females. Although mating is not essential for producing eggs, our study shows that even brief exposure to the semen and seminal fluids greatly enhances the oviposition and hatching efficiency, even if the mating occurs just before oviposition. However, those females mating before blood-feeding and those mating after blood-feeding produced 2266 and 2128 eggs, with hatching rates of 96.78% and 95.54%, respectively. Hence, the retention time of seminal fluid in the female seems to influence the number of eggs laid and the number of eggs hatched. CONCLUSIONS: In general, mating is possible in Ae. aegypti even minutes before oviposition and is sufficient to produce a greater number of viable eggs.

Prevalence and antimicrobial resistance of salmonellae isolated from eggs of local chicken in selected towns of Ethiopia.

BACKGROUND: Salmonellosis is one of the most common food-borne diseases in industrialised and developing countries. In recent year, an increase in antimicrobial resistance among different Salmonella serotypes has been observed. OBJECTIVE: A cross-sectional study was conducted to assess the prevalence and antimicrobial susceptibility of Salmonella isolated from local chicken eggs in four selected towns in Ethiopia. METHODS: A total of 115 eggs were examined to detect Salmonella by using standard microbiological methods. The susceptibilities of the isolates to nine antimicrobials were tested by the Kirby-Bauer disk diffusion method. RESULT: The study revealed that of the 115 eggs examined, 22 (19.1%) were positive for Salmonella of which 14 (12.2%) and 8 (7%) of the isolates were from shells and contents, respectively. The occurrence of Salmonella in egg shells and content and between different altitudes did not differ significantly (p > 0.05). Most isolates were resistant to more than three antimicrobials with a high resistance to kanamycin, ampicillin, nalidixic acid, cotrimoxazole, oxytetracycline and chloramphenicol. CONCLUSION: The results indicate the potential importance of local chicken eggs as source of multiple antimicrobial-resistant salmonellae and the need for proper cooking before consumption. Further studies are required to describe the epidemiology of Salmonella in various agroclimatic zones of Ethiopia.

Variability of pMGA/vlhA sequences among Mycoplasma gallisepticum field strains isolated from laying hens and their deformed eggs.

Mycoplasmosis, attributed to Mycoplasma gallisepticum, poses a significant challenge to poultry farming, leading to substantial economic losses and persistent infections within flocks. This bacterium harbours various surface proteins that are crucial for adhesion, transporter activity and evasion of the host immune response, facilitating its pathogenicity. One such key surface lipoprotein, referred to as pMGA or vlhA haemagglutinin, plays a pivotal role in adhesion processes. In this study, the clonal regions pMGA1.2 and pMGA1.3, as reported by Markham (M83178.1), were investigated to elucidate differences or similarities in the whole DNA sequences of M. gallisepticum field strains. The aim was to analyse sequence diversity within this region. Six internal primers were designed to amplify the target sequence, and isolates were obtained from both eggs and chickens sourced from laying hen flocks. Identification revealed 17 strains of M. gallisepticum and four strains of Mycoplasma synoviae, which were confirmed through the mgc2 and 16S rRNA genes, respectively. Positive and negative controls were established using the MGS6 and MSWUV1853 strains. Amplification results indicated a higher frequency of amplification proximal to the C-terminal region, with segments 4 (33.3 %) and 6 (27.8 %) being the most prevalent. Notably, none of the field strains exhibited the same amplification pattern as MGS6, and none of the strains characterized as M. synoviae amplified any primer set. Upon translation, the amino acid sequences from segments 4 and 6 were found to be compatible with conserved sequences within the Myco_haema protein domains of the genus Mycoplasma, specifically corresponding to Q7NAP3_MYCGA VlhA.3.04. The observed homology suggests a potential genetic transfer, while the variability identified in the pMGA or vlhA gene region of the field strains may have significant implications for protection against M. gallisepticum infection in chickens.

Text messages to improve young child diets: Results from a cluster-randomized controlled trial in Kanchanpur, Nepal.

The objective of this study was to test whether adding a text message campaign about the importance of eating eggs and other nutrition-related behaviours to an on-going package of large-scale, diverse social and behaviour change interventions would improve four types of nutrition-related knowledge and behaviour outcomes: child diets (egg consumption as the primary outcome), maternal diets, maternal nutrition knowledge, and maternal participation in additional interventions. The cluster-randomized controlled trial involved a repeat cross-sectional design, recruiting families with children 12-23 months of age at baseline and endline in one plains district of Nepal. Throughout the 1000-day period, 51 text messages were sent to each household at specific time points to reinforce ideal diets and other nutrition-related practices and promote engagement with community health workers and other intervention platforms. The primary outcome was egg consumption and dietary diversity among young children. We found no population-level effect. Some evidence supports that for those who received and opened the SMS intervention, it improved child egg consumption (odds ratio [OR]: 1.41, 95% confidence interval [CI]: 1.03-1.93), child minimum dietary diversity (OR: 1.36, 95% CI: 1.07-1.73), child dietary diversity scores (ß: 0.15, 95% CI: 0.01-0.24), as well as maternal IYCF knowledge (ß: 0.21, 95% CI: 0.08-0.35), participation in health mothers' group meetings (OR: 3.03, 95% CI: 1.91-4.84) and Bhanchhin Aama listenership (OR: 1.36, 95% CI: 1.07-1.73). This study highlights the importance of more research to understand the effectiveness of emerging digital interventions for behaviour change among specific populations, to facilitate nuanced targeting to those who can best benefit from these investments. Registered at clinicaltrials.gov with identifier NCT03926689.

Relationship between Village Chicken Availability and Dietary Diversity along a Rural-Urban Gradient.

Dietary diversity is one of the fundamental factors of nutritional security and a proxy used to measure diet quality. The objective of this study was to investigate the relationship between village chicken availability and the dietary diversity of households along a rural-urban gradient. Face-to-face interviews were conducted using a structured questionnaire in rural (n = 100), peri-urban (n = 100), and urban (n = 100) areas of Pietermaritzburg uMgungundlovu District, KwaZulu-Natal, in South Africa. A positive relationship between distance from the city center and village chicken flock sizes (p < 0.001) was observed. Consumption of vegetables increased with an increase in distance from the city center (p < 0.01). A quadratic relationship was observed between distance from the city center and consumption of livestock-derived foods (LDFs) (p < 0.05). Consumption of LDFs increased with an increase in village chicken flock sizes (p < 0.05). Consumption of vegetables increased with an increase in village chicken flock sizes (p < 0.01). Food variety score (FVS) increased with an increase in distance from the city center (p < 0.05). Assessing the availability of village chickens across rural-urban gradients is a worthy opportunity to utilize to improve households' dietary diversity and alleviate poverty. It can be concluded that expanding village flock sizes could enhance the dietary diversity of households.

ASCARIS SUUM EGG RECOVERY FROM SLUDGE SAMPLES AFTER PHASE EXTRACTION.

Some helminth test methods for sanitation samples include a phase extraction step to reduce lipid content and final pellet size before microscopy. Hydrophilic and lipophilic solutions are used to create 2 phases, with a plug of organic material or debris in between, whilst eggs are supposedly compacted at the bottom of the test tube. We tested 10% formalin, acetoacetic buffer, and acid alcohol as the hydrophilic solutions, and ethyl acetate and diethyl ether as the lipophilic solvents for egg recoverability from water, primary sludge, and fatty sludge. Normally, the supernatant and debris plug are discarded and the sedimented pellet of eggs is microscopically examined. We, however, also collected the entire supernatant plus debris plug to determine where eggs were possibly lost. We found that eggs were lost when samples were extracted with 10% formalin + ethyl acetate, 10% formalin + diethyl ether, acetoacetic buffer + ethyl acetate, and acetoacetic buffer + diethyl ether combinations (<50% egg recovery). Acid alcohol + ethyl acetate resulted in 93.2, 89.8, and 57.3% egg recovery in the pellet of water, primary sludge, and fatty sludge, respectively; however, the size of the final pellet was not reduced, defeating the purpose of the extraction step. We thus recommend that this step be excluded.

The impact of electromagnetic fields generated by high-voltage power lines on the spatial arrangement of pike (Esox Lucius Linnaeus 1758) embryos.

Urbanization and technological advancements result in the dispersion of antropogenic electromagnetic fields (EMF) that can affect on ecosystems. Therefore, it is important to understand their impact on the environment. Aquatic ecosystems are subject to EMF as part of various electricity sources, e.g., high-voltage transmission lines (HVTL). We examined the impact of EMF generated by HVTL on the spatial arrangement and survival of pike (Esox lucius) embryos. Fertilized eggs were incubated under two HVTL configurations 110 kV and 220 kV compared with a control group devoid of anthropogenic EMF. Embryo orientation and survival were monitored until blastopore closure. The control group showed dominance in the arrangement of embryos along the N-S, NNW-SSE, and NNE-SSW axes, with a slight prevalence of northern directions. EMF originating from HVTL did not exert a significant influence on the spatial arrangement of pike embryos, although some deviations from the arrangement noticed in the control group were observed. Increased embryo mortality was observed only at 110 kV site, but probably due to factors unrelated to EMF. In conclusion, EMF generated by HVTL did not significantly change pike embryo orientation or chances of survival. However, longer exposure or higher EMF levels could provoke notable reactions, requiring ongoing evaluation as power networks continue to spread more widely.

Influence of rosehip (Rosa canina L.) leaves as feed additive during first stage of laying hens on performances and egg quality characteristics.

This study aimed to evaluate the effects of rosehip leaves (RL) supplementation on the production performances and egg quality of laying hens during the first stage of laying. For that, 90 Lohmann Brown Classic hens, aged 26 wk, were randomly assigned to 3 dietary treatments as follows: a control diet (RL0), an experimental diet containing 0.5% rosehip leaves (RL0.5), and an experimental diet containing 1% rosehip leaves (RL1). The laying hens were accommodated in an experimental hall with identical pens (3.96 m2), and controlled microclimate conditions. The dietary treatments were administered for a 6-wk experimental period (26-32 d of age). During this time, production performance of hens was recorded. To assess the impact of RL on egg weight and its components evolution, egg yolk and albumen color, and primary chemical composition of egg constituents were determined every 2 wk. The effect on egg yolk fatty acids profile was determined only at the end of the trial. The results showed that RL0.5 and RL1 dietary treatments significantly improved laying rate, feed conversion ratio and egg mass compared with RL0 group. A significant effect on egg weight and its components was observed only over time, showing that RL has no effect on these parameters. Significant changes in egg yolk coloration were observed over time, with significant increases in L*, a* and b* chromomeric parameters in both RL0.5 and RL1 compared to RL0. Significant alterations were obtained from the sum of egg yolk fatty acids. The RL1 egg yolk presented significantly higher n-3, and significantly lower n-6 fatty acids, compared with both RL0 and RL0.5. Further research is warranted to explore the long-term effects of rosehip leaf supplementation on laying hens performance and egg quality.

Non-destructive prediction of fertility and sex in chicken eggs using the short wave near-infrared region.

The objective of this study was to evaluate the ability of a handheld near-infrared device (900-1600 nm) to predict fertility and sex (male and female) traits in-ovo. The NIR reflectance spectra of the egg samples were collected on days 0, 7, 14 and 18 of incubation and the data was analysed using principal component analysis (PCA), linear discriminant analysis (LDA) and support vector machines classification (SVM). The overall classification rates for the prediction of fertile and infertile egg samples ranged from 73 % to 84 % and between 93 % to 95 % using LDA and SVM classification, respectively. The highest classification rate was obtained on day 7 of incubation. The classification between male and female embryos achieved lower classification rates, between 62 % and 68 % using LDA and SVM classification, respectively. Although the classification rates for in-ovo sexing obtained in this study are higher than those obtained by chance (50 %), the classification results are currently not sufficient for industrial in-ovo sexing of chicken eggs. These results demonstrated that short wavelengths in the NIR range may be useful to distinguish between fertile and infertile egg samples at days 7 and 14 during incubation.

Chemical threats for the sentinel Pygoscelis adeliae from the Ross Sea (Antarctica): Occurrence and levels of persistent organic pollutants (POPs), perfluoroalkyl substances (PFAS) and mercury within the largest marine protected area worldwide.

The Ross Sea Marine Protected Area (RS-MPA) hosts endemic species that have to cope with multiple threats, including chemical contamination. Adèlie penguin is considered a good sentinel species for monitoring pollutants. Here, 23 unhatched eggs, collected from three colonies along the Ross Sea coasts, were analysed to provide updated results on legacy pollutants and establish a baseline for newer ones. Average sum of polychlorinated biphenyls (∑PCBs) at the three colonies ranged 20.9-24.3 ng/g lipid weight (lw) and included PCBs IUPAC nos. 28, 118, 153, 138, 180. PCBs were dominated by hexachlorinated congeners as previously reported. Hexachlorobenzene (HCB) and p,p'-dichlorodiphenyldichloroethylene (p,p'-DDE) ranged between 134 and 166 and 181-228 ng/g lw, respectively. Overall, ∑PCBs was exceeded by pesticides, contrary to previous studies from the Ross Sea. Sum of polybrominated diphenyl ethers (∑PBDEs) ranged between 0.90 and 1.18 ng/g lw and consisted of BDE-47 (that prevailed as expected, representing 60-80 % of the ∑PBDEs) and BDE-85. Sum of perfluoroalkyl substances (∑PFAS) ranged from 1.04 to 1.53 ng/g wet weight and comprised five long-chain perfluorinated carboxylic acids (PFCAs), perfluorooctane sulfonate (PFOS), perfluorohexane sulfonate (PFHxS) and perfluorooctanoic acid (PFOA); perfluorooctane sulfonamide (PFOSA) was also detected. The PFAS profile was dominated by PFCAs as already observed in Arctic seabirds. Mercury ranged from 0.07 to 0.15 mg/kg dry weight similarly to previous studies. Legacy pollutants confirmed their ongoing presence in Antarctic biota and their levels seemed mostly in line with the past, but with minor variations in some cases, likely due to continued input or release from past reservoirs. PFAS were reported for the first time in penguins from the Ross Sea, highlighting their ubiquity. Although further studies would be useful to increase the sample size and accordingly improve our knowledge on spatial and temporal trends, this study provides interesting data for future monitoring programs within the RS-MPA that will be crucial to test its effectiveness against human impacts.

Bufadienolide-Fatty Acid Conjugates from the Fertilized Eggs of Toad Bufo gargarizans: Isolation, Characterization, Toxicity, and Antiproliferative Evaluation.

Bufadienolides (BDs) are a class of naturally occurring toxins present in amphibian toads. Serving as the chemical weapons, they exist not only in the adult toads but also in toad eggs. Guided by mass spectrometry (MS)-based component analysis and feature-based molecular networking (FBMN), 30 bufadienolide-fatty acid conjugates (BDFs) were isolated from the fertilized eggs of toad Bufo gargrizans, including 25 previously undescribed compounds (1-25). Their chemical structures were elucidated by extensive spectroscopic analysis, chemical methods, and GC-MS. The toxicities of all BDFs and their corresponding free BDs were assessed using the zebrafish model. The structure-toxicity relationship analysis showed that the modification of BDs by hydroxy fatty acids can cause a significant increase of the toxicity. Furthermore, all the isolated compounds were evaluated for their antiproliferative activities in pancreatic cancer cell lines ASPC-1 and PANC10.05. The structure-activity relationship (SAR) analysis revealed that BDFs with hellebrigenin as the bufogenin moiety (6 and 7) exhibited the most potent antiproliferative effect. Further investigation into their functional mechanism demonstrated that 6 and 7 induced apoptosis in pancreatic cancer cells PANC10.05 and significantly suppressed the expression of the apoptosis-related gene c-MYC. In addition, 6 and 7 effectively inhibited the expression of the PI3K/Akt/mTOR pathway in PANC10.05. Moreover, we assessed the efficacy of 6 and 7 on cancer cells from various tissues and observed their broad-spectrum antiproliferative activity.

Diagnosis of Human Trematode Infections.

Digenetic trematodes form a major group of human parasites, affecting a large number of humans, especially in endemic foci. Over 100 species have been reported infecting humans, including blood, lung, liver and intestinal parasites. Traditionally, trematode infections have been diagnosed by parasitological methods based on the detection and the identification of eggs in different clinical samples. However, this is complicated due to the morphological similarity between eggs of different trematode species and other factors such as lack of sensitivity or ectopic locations of the parasites. Moreover, the problem is currently aggravated by migratory flows, international travel, international trade of foods and changes in alimentary habits. Although efforts have been made for the development of immunological and molecular techniques, the detection of eggs through parasitological techniques remains as the gold standard for the diagnosis of trematodiases. In the present chapter, we review the current status of knowledge on diagnostic techniques used when examining feces, urine, and sputum and also analyze the most relevant characteristics used to identify eggs with a quick key for the identification of eggs.

Evaluation of the Quality of Guinea Fowl (Numida meleagris) Eggs from Free-Range Farming Depending on the Storage Period and Age of Laying Hens.

The aim of the study was to determine the changes occurring in the eggs of helmeted guinea fowl (Numida meleagris) from free-range farming in relation to the laying season and storage time. The experimental material consisted of 360 guinea fowl eggs, collected in the first, second and third laying seasons and stored for 7, 14 and 21 days. After each period, physical and physicochemical characteristics of the eggs were determined, as well as the basic chemical composition and mineral content of the albumen and yolk and the yolk fatty acid profile. The age of the guinea fowls affected certain physical parameters of the eggs. The egg weight, shape index and shell thickness increased with the age of the laying hens; however, a decrease in the proportion of shell in the egg was demonstrated. Storage time had a significant effect on egg weight, weight loss during storage and air cell height. Significant differences were found in the chemical composition of guinea fowl eggs depending on the age of the laying hens. Eggs obtained from older laying hens were characterized by higher yolk fat content and lower ash content, while the albumen contained higher water content and lower ash content. During the three-year laying period, changes were observed in the mineral composition of the eggs. The fatty acid profile underwent significant changes; however, no important differences were observed in the total content of SFA, MUFA, PUFA and n-6 fatty acids. Conversely, significant differences were found for n-3 acids and the n-6/n-3 ratio. Eggs in the first and second laying seasons exhibited the most favorable composition. The slow dynamics of changes occurring in successive laying seasons and egg storage time indicated that the raw material studied was safe and could be used by consumers.

Chickens' eggs and the livers of farm animals as sources of perfluoroalkyl substances.

Introduction: This study focuses on perfluoroalkyl substance (PFAS) content in chickens' eggs and the livers of farm animals. Material and Methods: Chickens' eggs (n = 25) and the livers of cows (n = 10), chickens (n = 7) and horses (n = 3) were collected from various regions of Poland. Samples were analysed using the isotope dilution technique with liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS). Results: The mean lower bound (LB) sum of four PFAS (∑4 PFAS) concentrations (perfluorooctanesulfonic acid (PFOS), perfluorooctanoic acid (PFOA), perfluorononanoic acid (PFNA) and perfluorohexanesulfonic acid (PFHxS)) were the highest in cows' livers (0.52 µg/kg) and much lower in chickens' (0.17 µg/kg) and horses' livers (0.13 µg/kg) and chickens' eggs (0.096 µg/kg). The ratio of ∑4 PFASs to the limits set by Commission Regulation (EU) 2023/915 was <7% for liver and <6% for eggs. Linear PFOS was the compound with the highest detection frequency (8% in eggs and 48% in all livers). In cows' livers it was detected in 80% of samples. The estimated exposure to LB ∑4 PFASs via consumption of liver tissue from farm animals (assuming 50 g and 100 g portions) was <52% of the tolerable weekly intake (TWI) for children and <17% of the TWI for adults. Dietary intake via the average portion of three eggs led to low exposure of <15% for children and <5% for adults. Conclusion: Neither eggs nor the livers of chickens or horses as analysed in this study are significant sources of PFASs, while cows' livers might contribute significantly to a child's overall dietary intake. Further investigation of PFOS in farm animal livers should be conducted.

Different narcotic gases and concentrations for immobilization of ostrich embryos for in-ovo imaging.

In-ovo imaging using avian eggs has been described as a potential alternative to animal testing using rodents. However, imaging studies are hampered by embryonal motion producing artifacts. This study aims at systematically comparing isoflurane, desflurane and sevoflurane in three different concentrations in ostrich embryos. Biomagnetic signals of ostrich embryos were recorded analyzing cardiac action and motion. Ten groups comprising eight ostrich embryos each were investigated: Control, isoflurane (2%, 4%, and 6%), desflurane (6%, 12%, and 18%) and sevoflurane (3%, 5%, and 8%). Each ostrich egg was exposed to the same narcotic gas and concentration on development day (DD) 31 and 34. Narcotic gas exposure was upheld for 90 min and embryos were monitored for additional 75 min. Toxicity was evaluated by verifying embryo viability 24 h after the experiments. Initial heart rate of mean 148 beats/min (DD 31) and 136 beats/min (DD 34) decreased over time by 44-48 beats/minute. No significant differences were observed between groups. All narcotic gases led to distinct movement reduction after mean 8 min. Embryos exposed to desflurane 6% showed residual movements. Isoflurane 6% and sevoflurane 8% produced motion-free time intervals of mean 70 min after discontinuation of narcotic gas exposure. Only one embryo death occurred after narcotic gas exposure with desflurane 6%. This study shows that isoflurane, desflurane and sevoflurane are suitable for ostrich embryo immobilization, which is a prerequisite for motion-artifact free imaging. Application of isoflurane 6% and sevoflurane 8% is a) safe as no embryonal deaths occurred after exposure and b) effective as immobilization was observed for approx. 70 min after the end of narcotic gas exposure. These results should be interpreted with caution regarding transferability to other avian species as differences in embryo size and incubation duration exist.

Occurrence, antimicrobial resistance, and molecular characterization of Salmonella enterica from chicken products and human in Wasit Governorate of Iraq.

Background: Salmonella infections are considered the most common foodborne pathogens responsible for zoonotic infections and food poisoning in humans and animal species such as birds. Antimicrobial resistance is considered a global anxiety because it causes human public health repercussions, as well as leads to an increase in animal morbidity and death. Aim: The aims of this study are the isolation and identification of Salmonella enterica, as well as to investigate the antimicrobial susceptibility test (AST) and the molecular characteristics using polymerase chain reaction (PCR) and sequences for isolates from chicken products (eggs, livers, and minced meat) and human in the Wasit Governorate of Iraq. Methods: A total of 300 samples (150 chicken product samples including eggs, livers, and minced meat, and 150 human fecal samples) were collected from the Wasit governorate of Iraq from January to December 2022. The bacterial isolation was done according to recommendations of ISO 6579 standard and the Global Foodborne Infections Network laboratory protocol. Serotyping test and AST were done by using 19 antibiotic agents according to the recommendations of the Clinical and Laboratory Standards Institute, 2022 by using disc diffusion susceptibility test and Vitik 2 test. Finally, the suspected isolates were confirmed using the conventional PCR method and sequencing for a unique rRNA gene. Results: The results showed that the isolation percentage of S. enterica in chicken products was 8.66% (12% eggs, 6% livers, and 8% minced meat), while in humans it was 4.6%. Also, showed 100% of Salmonella typhi in humans. While, in chicken eggs S. typhi, Salmonella typhimurium, and Salmonella enteritidis were 50%, 33.33%, and 16.66%, respectively. Also, showed 100% of S. typhimurium in both livers and minced meat. The AST in human isolates showed resistance to Ampicillin, Cefotaxime, Ceftazidime, Cefepime, Amikacin, Gentamicin, Ciprofloxacin, Norfloxacin, and Ceftriaxone, while no resistance to Amoxicillin, Pipracillin, Ertapenem, Imipenem, Meropenem, Fosfomycin, Nitrofurantoin, Trimethoprim, Azithromycin, and Tetracycline. In chicken products, isolates were resistant with different percentages to Amikacin, Gentamicin, Tetracycline, Ciprofloxacin, Norfloxacin, Nitrofurantoin, Ampicillin, Cefotaxime, Ceftazidime, Cefepime, and Trimethoprim; while no resistance to Amoxicillin, Pipracillin, Ertapenem, Imipenem, Meropenem, Fosfomycin, Azithromycin, and Ceftriaxone. Sequencing by using rRNA gene was done for four PCR products. Conclusion: This study showed the presence of genetic mutations for S. enterica which led to variations in the molecular characteristics, and antimicrobial drug resistance of S. enterica isolated from chicken products and humans.