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Beta-attenuation monitors are used worldwide to monitor PM mass concentration with high temporal resolution. Hourly PM10 and PM2. 5 dry mass concentrations are publicly available with the tacit assumption that water is effectively removed prior to the measurement. However, as both the filter material of the monitor and the aerosol particles are capable of retaining a significant amount of water even at low relative humidities, the basic assumption may not be valid, resulting in significant bias in reported PM10 and PM2. 5 concentrations. Here we show that in PM10 measurement, particle-free air can produce apparent hourly average PM concentrations in the range of −13–+21 µg m−3 under conditions of fluctuating relative humidity. Positive and negative apparent readings are observed with increasing and decreasing relative humidities, respectively. Similar phenomena have been observed when the instrument filter was previously loaded with atmospheric aerosol. As a result the potential measurement biases in hourly readings arising from the interaction with water may be in the range of −53… + 69 %.

Secondary aerosol particle production via new particle formation (NPF) has been shown to be a major contributor to the global aerosol load. NPF has also been observed frequently in urban environments. Here, we investigate the effect of regional NPF on urban aerosol load under well-defined atmospheric conditions. The Carpathian Basin, the largest orogenic basin in Europe, represents an excellent opportunity for exploring these interactions. Based on long-term observations, we revealed that NPF seen in a central large city of the basin (Budapest) and its regional background occur in a consistent and spatially coherent way as a result of a joint atmospheric phenomenon taking place on large horizontal scales. We found that NPF events at the urban site are usually delayed by > 1 h relative to the rural site or even inhibited above a critical condensational sink level. The urban processes require higher formation rates and growth rates to be realized, by mean factors of 2 and 1.6, respectively, than the regional events. Regional- and urban-type NPF events sometimes occur jointly with multiple onsets, while they often exhibit dynamic and timing properties which are different for these two event types.

The third intensive measurement period (IMP) organised by the European Monitoring and Evaluation Programme (EMEP) under the UNECE CLTRAP took place in summer 2012 and winter 2013, with PM10 filter samples concurrently collected at 20 (16 EMEP) regional background sites across Europe for subsequent analysis of their mineral dust content. All samples were analysed by the same or a comparable methodology. Higher PM10 mineral dust loadings were observed at most sites in summer (0.5–10 µg m−3) compared to winter (0.2–2 µg m−3), with the most elevated concentrations in the southern- and easternmost countries, accounting for 20–40 % of PM10. Saharan dust outbreaks were responsible for the high summer dust loadings at western and central European sites, whereas regional or local sources explained the elevated concentrations observed at eastern sites. The eastern Mediterranean sites experienced elevated levels due to African dust outbreaks during both summer and winter. The mineral dust composition varied more in winter than in summer, with a higher relative contribution of anthropogenic dust during the former period. A relatively high contribution of K from non-mineral and non-sea-salt sources, such as biomass burning, was evident in winter at some of the central and eastern European sites. The spatial distribution of some components and metals reveals the influence of specific anthropogenic sources on a regional scale: shipping emissions (V, Ni, and SO42−) in the Mediterranean region, metallurgy (Cr, Ni, and Mn) in central and eastern Europe, high temperature processes (As, Pb, and SO42−) in eastern countries, and traffic (Cu) at sites affected by emissions from nearby cities.

Absorption photometers for real time application have been available since the 1980s, but the use of filter-based instruments to derive information on aerosol properties (absorption coefficient and black carbon, BC) is still a matter of debate. Several workshops have been conducted to investigate the performance of individual instruments over the intervening years. Two workshops with large sets of aerosol absorption photometers were conducted in 2005 and 2007. The data from these instruments were corrected using existing methods before further analysis. The inter-comparison shows a large variation between the responses to absorbing aerosol particles for different types of instruments. The unit to unit variability between instruments can be up to 30% for Particle Soot Absorption Photometers (PSAPs) and Aethalometers. Multi Angle Absorption Photometers (MAAPs) showed a variability of less than 5%. Reasons for the high variability were identified to be variations in sample flow and spot size. It was observed that different flow rates influence system performance with respect to response to absorption and instrumental noise. Measurements with non absorbing particles showed that the current corrections of a cross sensitivity to particle scattering are not sufficient. Remaining cross sensitivities were found to be a function of the total particle load on the filter. The large variation between the response to absorbing aerosol particles for different types of instruments indicates that current correction functions for absorption photometers are not adequate.

In contrast to most angiosperms, Torenia contains a naked embryo sac and therefore has been considered since many years as an exciting model plant to study the double fertilization process of flowering seed plants. It is thus not surprising that the isolation of protoplasts from the female gametophyte has been reported already 20 years ago by Mol, the isolation of megaspores and megagametophytes has been published by the authors of this manuscript in 1996 and in 1999. The isolation of the male gametophyte and of sperm cells was published by the authors in 2004. The isolation of viable Torenia sperm cells is a crucial part of the elaboration of an in vitro fertilization system. Torenia sperm cells were isolated from in vivo — in vitro cultured pollen tubes. In this system pollen tubes first grow inside a cut style then follow their elongation in a solid isolation medium. The medium contained agarose in order to detain pollen tube contents. Released sperm cells and enzymatically isolated egg cells were collected and handled using glass micropipettes and transmitted to an electrofusion apparatus or polyethylene glycol containing media for fusion probes.

Game meat is derived from non-domesticated, free-ranging wild animals and plays an important role in human nutrition, but it is recognized as a source of food-borne and drug-resistant pathogens impacting food safety. The present review aimed to provide a comprehensive analysis of the frequency of isolation and antimicrobial resistance (AMR) profiles of major foodborne pathogens from the Enterobacteriaceae , including Salmonella , Escherichia , and Yersinia genera, in wild ungulates, across Europe in the 21st century. A systematic search was conducted via the Google Scholar database using the PRISMA guidelines. In this regard, the content of a total of 52 relevant scientific publications from both European Union (n = 10) and non-European Union countries (n = 3) was processed, highlighting the main scientific achievements and indicating knowledge gaps and future perspectives. The studies highlighted that Salmonella spp. was the most commonly encountered pathogen, and significant AMR levels were noticed for the isolated strains, especially against penicillin (32.8%) and amoxicillin (32.1%). This review underscores the importance of monitoring the presence of food-borne pathogens and their AMR in wildlife as important public health and food safety concerns.

The present study investigates the prevalence and etiology of subclinical mastitis in Țurcana sheep flocks located in south-western Romania. Milchtest and California Mastitis Test (CMT), were used for the detection of subclinical mastitis. A number of 360 milk samples across three lactation phases were analyzed. Subclinical mastitis was identified in 146 (40.6%) of sheep milk samples. Only milk samples (n = 146) collected from sheep diagnosed with subclinical mastitis were considered for microbiological analyses. Thus, Staphylococcus aureus (129/146; 88.4%) was identified as the predominant pathogen followed by Streptococcus spp. (14/146; 9.6%) Enterococcus spp. (7/146; 4.8%), Pseudomonas aeruginosa (6/146; 4.1%) and Klebsiella pneumoniae (5/146; 3.4%). Antimicrobial susceptibility of isolated strains of Staphylococcus aureus, Streptococcus spp. and Enterococcus spp. was also tested. The results showed varying resistance patterns depending on the farm and microorganism. In heard A the highest resistance of Staphylococcus aureus was identified to polymyxin B (79.7%) followed by novobiocin (73.4%) and tetracycline (45.3%); Streptococcus spp. to streptomycin (100%), novobiocin (100%), tetracycline (100%) and Enterococcus spp. to tylosin (100%), streptomycin (75.0%) and tetracycline (75.0%). In herd B, Staphylococcus aureus showed high resistance rates to polymyxin B (86.3%), novobiocin (74.5%) and cloxacillin (39.2%); Streptococcus spp. strains to streptomycin (100%), novobiocin (100%), tetracycline (100%) and Enterococcus spp. to gentamicin (100%), streptomycin (66.7%) and erythromycin (66.7%). The obtained preliminary results indicate a potential risk that future treatments of sheep mastitis may not be effective, and demonstrated that raw unpasteurized milk can harbor antimicrobial resistance pathogens posing a threat to public health. However, further investigations involving molecular research on resistance genes are required to draw comprehensive conclusions., Competing Interests: Declarations. Ethical approval: Even though the research entitled “Etiology and Antimicrobial Resistance of Subclinical Mastitis Pathogens Staphylococcus aureus, Streptococcus spp. and Enterococcus spp. in Sheep Milk” included live animals (ovines), however, they were not directly affected or used because the samples processed and analyzed in the study were represented by milk samples collected from sheep during milking stages. The statement was included in the Bioethics Commission Approval No. 5639. The experiments assure the proper accommodation and animal handling in accordance with the Directive 2010/63/EU. Also the experimental protocol includes methods for preventing and reducing animal distress in accordance to the recommendation of European Union and Romanian Legislation. Competing interests: The authors declare no competing interests., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

This study investigated the bacteriological quality of raw ovine milk produced by farms located in the Banat region of Romania. Additionally, the pathogenic potential and antimicrobial resistance of the isolated Escherichia coli strains were evaluated. A total of 95.8% (69/72) of the screened bulk tank milk samples, collected at the farm level immediately after milking, demonstrated appropriate total aerobic mesophilic bacteria (TMB) counts, varying from 3.32 to 6.09 log10 CFU/mL. However, 4.2% (3/72) of the samples were above the regulatory limit of 6.18 log10 CFU/mL. E. coli was identified in 66.6% of the examined samples, and from the total number ( n = 48) of isolates, 18.8% harbored the stx 2 gene, highlighting pathogenic potential. Antimicrobial susceptibility testing with the Vitek2 system of the isolated E. coli strains revealed resistance against ampicillin (45.8%), gentamicin (20.8%), ticarcillin-clavulanic acid (18.8%), cephalexin (18.8%), amoxicillin-clavulanic acid (8.3%), and trimethoprim-sulfamethoxazole (2.1%). Additionally, 64.6% of the strains showed intermediate resistance against amoxicillin-clavulanic acid, while no resistance was recorded against imipenem. Five (18.5%) strains were multidrug-resistant. This study's results underline hygienic sanitary deficiencies during the milking phase and indicate that raw ovine milk can be contaminated with pathogenic and multidrug-resistant E. coli strains, highlighting a potential risk to public health. Further studies are encouraged to better understand the risk posed to the consumer via the consumption of ovine milk and derived products.

Poultry carcasses may be reservoirs for the zoonotic transmission of antimicrobial-resistant bacteria to humans and pose a major public health hazard. During the isolation of Salmonella from poultry and other foods, many of the presumptive typical Salmonella colonies on xylose lysine deoxycholate (XLD) agar were found to lack the invA gene, which is the specific target gene for Salmonella spp. Therefore, the current study aimed to estimate the prevalence and antimicrobial resistance profiles of extensively drug-resistant invA -negative non- Salmonella isolates recovered from native Egyptian chicken carcasses as presumptive Salmonella colonies on XLD agar. The non- Salmonella isolates were detected in 84% (126/150) of the examined native Egyptian chicken carcasses and classified into five genera, with prevalence rates of 64% (96/150), 14% (21/150), 6.7% (10/150), 3.3% (5/150), and 1.3% (2/150) for Proteus , Citrobacter , Shigella , Pseudomonas , and Edwardsiella , respectively. One hundred and ninety-five invA -negative, non-verified presumptive Salmonella isolates were recovered and classified at the species level into Proteus mirabilis (132/195; 67.7%), Proteus vulgaris (11/195; 5.6%), Citrobacter freundii (26/195; 13.3%), Shigella flexneri (8/195; 4.1%), Shigella sonnei (6/195; 3.1%), Shigella dysenteriae (3/195; 1.5%), Pseudomonas fluorescens (6/195; 3.1%), and Edwardsiella tarda (3/195; 1.5%). All (195/195; 100%) of these isolates showed resistance against cefaclor and fosfomycin. Additionally, these isolates showed high resistance rates of 98%, 92.8%, 89.7%, 89.2%, 89.2%, 86.7%, 80%, 78.5%, 74.4%, and 73.9% against cephalothin, azithromycin, vancomycin, nalidixic acid, tetracycline, sulfamethoxazole/trimethoprim, cefepime, gentamicin, cefotaxime, and ciprofloxacin, respectively. Interestingly, all (195/195; 100%) of the identified isolates were resistant to at least five antibiotics and exhibited an average MAR (multiple antibiotic resistance) index of 0.783. Furthermore, 73.9% of the examined isolates were classified as extensively drug-resistant, with an MAR index equal to 0.830. The high prevalence of extensively drug-resistant foodborne Proteus , Citrobacter , Shigella , Pseudomonas , and Edwardsiella isolated from native chicken carcasses poses a great hazard to public health and necessitates more monitoring and concern about the overuse and misuse of antibiotics in humans and animals. This study also recommends the strict implementation of GHP (good hygienic practices) and GMP (good manufacturing practices) in the chicken meat supply chain to protect consumer health.

Campylobacter spp. is recognized as one of the most common pathogens involved in the development of gastrointestinal infections in humans. The current study aimed to enhance the knowledge on the occurrence and molecular characterization of Campylobacter spp. in pigs and cattle origin caecum samples (n = 56) collected in one year, from nine Romanian slaughterhouses, and to determine the antimicrobial resistance profile of the isolated strains. All Campylobacter spp. strains (n = 41) isolated from swine and cattle caecum samples were analyzed in terms of antimicrobial resistance, in accordance with the EURL protocol and with the Commission Implementing Decision No. 2020/1729. The prevalence rate for C. coli. in pig caecum samples was 92.3% (36/39), and the prevalence of C. jejuni, in cattle origin samples was 29.4% (5/17). C. coli strains isolated from pigs proved resistant to tetracycline 75% (27/36), ciprofloxacin 69.4% (25/36), erythromycin 8.3% (3/36), ertapenem 2.8% (1/36) and gentamicin 2.8% (1/36), but no resistance was observed towards chloramphenicol. C. jejuni strains originating from cattle expressed resistance to ciprofloxacin 60.0% (3/5) and tetracycline 20.0% (1/5), but they were susceptible to chloramphenicol, erythromycin, ertapenem and gentamicin. In the present study, 19.5% (7/36) C. coli strains isolated from pigs were identified as multidrug-resistant (MDR) bacteria. The obtained results demonstrated that pigs especially, but cattle to, can be considered important natural reservoirs for zoonotic multidrug-resistant Campylobacter strains, having a stimulating effect for further studies aiming at the molecular screening of the genotypic antimicrobial resistance processing of a higher number of samples., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

Semidry beef luncheon may undergo deteriorative changes during storage at ambient temperatures in tropical and subtropical regions including Egypt. This study was conducted in a meat plant in Egypt with the aim of overcoming the economic losses from the returns of spoiled unsold beef luncheon displayed in grocery stores at adverse summer temperatures of 37 °C or more. Ten approaches were applied using different preservatives, comprising sodium nitrite, nisin, potassium sorbate, and organic acid salts (a combination of sodium lactate, sodium acetate, and sodium diacetate). In addition, the product was cooked at different temperatures and was stored for 21 days at 37 °C, during which time the shelf life, microbial quality, pH, and sensory attributes were investigated. By Day 21 of storage, the luncheon contained 50 mg/kg sodium nitrite, 25 mg/kg nisin, and 1000 mg/kg organic acid salts and, when cooked at a final core temperature of 92 °C, exhibited reductions in aerobic plate count, anaerobic plate count, lactic acid bacterial count, and mold and yeast counts by 4.32, 3.54, 3.47, and 1.89 log 10 CFU/g, respectively, when compared with the control. The sensory attributes and pH were also maintained in the final products of such treatment, with no product return and the avoidance of economic loss. This study presents a novel approach for solving the major problem of the deteriorative changes that occur in semidry luncheon sausage and similar meat products which require rejection with a huge economic loss, especially in tropical and semitropical areas of the world that have similar problems of high climatic temperatures and a low availability of energy or technological resources.

Meat discoloration, lipid oxidation, and undesirable texture are inevitable phenomena in basa fish fillets during storage, which in turn limits their exportation as well as decreases consumer acceptability. In addition, increasing consumers' requirements for high-quality, minimally processed, and ready-to-cook fish fillets with an extended shelf-life is a great challenge, particularly with lifestyle changes. Accordingly, this study aimed to improve the quality, lipid stability, fatty acid profile, and lipid nutritional quality indices (LNQI) of basa fish fillets during chilling storage at 4 °C for 15 days using pepsin enzyme (E, 0.1%), rosemary oil (R, 0.5%), citric acid (CA, 0.5%), and their combination (0.1% E + 0.5% R; 0.1% E + 0.5% CA; and 0.1% E + 0.5% R + 0.5% CA). Our results revealed that all treated samples exhibited a significant increase in protein content, a significant decrease in fat content, and a marked reduction in pH, total volatile base nitrogen (TVBN), thiobarbituric acid (TBA), free fatty acids, and shear force (SF) values in comparison to control ones. Moreover, significant improvements in sensory scores, color stability, fatty acid profile, LNQI, and microbial quality of all treated samples were observed. Such findings were more pronounced in samples treated with a mixture of pepsin, rosemary, and citric acid (TVBN: 2.04 vs. 6.52 mg%; TBA: 0.40 vs. 2.68 mg malonaldehyde/Kg; and SF: 8.58 vs. 19.51 Kgf). Based on the obtained results, there was an extension for the shelf life of all treated basa fish fillet samples, especially in samples treated with a mixture of pepsin, rosemary, and citric acids when compared with the control samples (˃15 days versus 10 days). Additionally, eucalyptol, camphor, isoborneol, and α-pinene are the main components of rosemary, with great antioxidant and antimicrobial activity. In conclusion, the mixture of pepsin, rosemary, and citric acid can be applied easily in the seafood industry and at the household level to provide ready-to-cook fish fillets of high quality with great health benefits.

The study was undertaken to investigate the main causes of carcass and organ condemnations, as well as to estimate the financial losses suffered by a cattle slaughterhouse. In this regard, an active abattoir survey, based on standard post-mortem inspection procedures for meat, was conducted on 151,741 cattle, from January 2021 to December 2022. Overall, 13.27% ( n = 20,125) of the carcasses expressed lesions or pathological conditions and, out of them, 1.15% ( n = 1738) were totally confiscated, while another 12.12% ( n = 18,387) were partially admitted for human consumption. In the case of organs, the general inspection data reveal that 12.28% ( n = 18,630), 7.56% ( n = 11,477), 1.89% ( n = 2862), and 0.27% ( n = 412) of the examined liver, lung, heart, and kidney specimens presented one or more types of abnormalities. In addition, regarding the types of specific pathological findings, dystrophies/anomalies (69.8%), circulatory disorders (40.6%), fecal contamination (60.9%), and suspected bacterial/viral infections showed a dominant occurrence in the liver, lung, heart, and kidneys, respectively. Consequently, the total direct financial losses resulting from edible part condemnation over the two years was estimated at EUR 4,021,717.3, which represents 1.17% of the total achievable net revenue without carcass and organ condemnation. Of this, EUR 3,661,400.4 (1.07%) and EUR 360,316.9 (8.73%) was related to carcass and organ condemnation, respectively. The study results demonstrate that the post-mortem inspection of meat at the slaughterhouse level plays a crucial role in identifying pathological lesions, in addition to some other issues, such as fecal contamination or non-compliant laboratory results, relevant to both public health and economic factors.

Rumen flukes are geographically widespread trematodes affecting wild and domestic ruminants. The juvenile forms, which are found in the small intestine, are more pathogenic compared to the adults. Severe diarrhoea and weight loss are the major clinical signs, and the disease might be fatal in severely infested individuals. In the last decade, paramphistomosis has been described as an emerging parasitic disease in Europe. This study aimed to identify the rumen fluke species in wild ruminants from western Romania. Fifty-two pre-stomachs obtained from roe deer ( Capreolus capreolus ) that were hunted on 14 hunting grounds from Timiș and Arad counties were examined for the presence of paramphistomes. Three (9.09%) out of 33 samples were positive in Timiș County, and one (5.26%) out of 19 samples was positive in Arad County. Subsequent PCR testing revealed that three samples were positive for Calicophoron daubneyi and one for Paramphistomum leydeni . The presence of C. daubneyi and P. leydeni in roe deer has not been previously reported in Romania. Two Paramphistomum species- C. daubneyi and P. leydeni -were revealed as the main species of rumen flukes in roe deer from forests in Romania.

The aim of this study was to assess the hygiene of pork, beef, and poultry carcasses and to determine the phenotypic antibiotic susceptibility of the bacteria embedded in the biofilm formed on the carcasses kept in cooling chambers for at least three days. The level of hygiene was assessed by determining the total aerobic colony count (TACC) and the Enterobacteriaceae level in different sampling points of the carcasses, along with the detection of E. coli and Pseudomonas spp. embedded in the biofilm. Furthermore, the E. coli and Pseudomonas spp. isolates were tested for antimicrobial resistance profiles. A total of 130 samples collected from pork, beef, and poultry from processing units were analyzed to determine the total aerobic colony count as well as to measure the level of Enterobacteriaceae found on the carcasses. The antimicrobial susceptibility of 44 Escherichia coli and eight Pseudomonas spp. strains isolated from the carcasses were assessed using the Vitek 2 system using two different cards. Overall, the regulatory limits for the TACC were exceeded in 7.6% of the samples, and 65% of the samples exceeded the regulatory limits for Enterobacteriaceae levels. The antimicrobial susceptibility tests of the E. coli isolates analyzed with the AST-GN27 card revealed the highest resistance to be that towards ampicillin (76.1%), followed by cefazolin (71.4%), amoxicillin/clavulanic acid (61.9%), nitrofurantoin (52.3%), cefoxitin (47.6%), tetracycline (38.1%), piperacillin, norfloxacin (19%), trimethoprim-sulfamethoxazole (11.9%), cefotaxime (9.5%), ceftazidime, cefazolin, amikacin, gentamicin, and ciprofloxacin (4.7%). However, all of the isolates were sensitive to piperacillin-tazobactam and imipenem. Thirty-two (61.5%; 95% CI 47.9-73.5) out of fifty-two isolates exhibited multidrug resistance, resulting in the expression of 10 resistance profiles. The findings of this study highlight serious hygienic and sanitary deficiencies within the meat processing units and demonstrate that the resulting meat can harbor Multidrug-resistant Escherichia coli and Pseudomonas spp., both of which pose a serious public health risk. However, further research with a larger number of samples is required to reach thorough results.

The study was designed to evaluate the effects of the total alkaloid extract of Algerian Peganum harmala seeds on sexual behavior and male reproductive function. After two weeks of acclimatization, the male mice were randomly divided into four groups (seven mice in each group). For 35 days, the extract was administered orally at dose levels of 6.25, 12.5, and 25 mg/kg body weight per day to the respective groups of male mice (n = 7) and normal saline daily to the control group. On day 28, sexual behavior parameters were recorded. At the end of the trial, reproductive organ weights, sperm quality, seminal fructose, and testosterone hormone levels were evaluated. The three treated groups were compared with the control using statistical variance analysis (one-way ANOVA, p < 0.05), followed by Tukey's test. The results of the groups treated with 12.5 and 6.25 mg/kg of P. harmala alkaloid revealed the MF and IF parameters to be the lowest compared to the control group ( p < 0.05). However, the male mice treated with 25 mg/kg recorded the highest values. A low significant value of ML was observed in the group treated with 25 mg/kg of the total alkaloid extract of P. harmala compared to the control group ( p < 0.01), while a rise was observed in the concentration group treated with 6.25 mg/kg. Regarding IL, the male mice treated with different concentrations of the total alkaloid extract of P. harmala recorded a higher time than the control group. Moreover, an increase in the gonadosomatic index was noticed in all groups compared to the control group. However, there was a significant ( p < 0.01) decrease in the sperm counts of the groups treated with 12.5 mg/kg and 6.25 mg/kg. However, there was no significant difference in the motility, membrane integrity, and total antioxidant capacity of sperm cells compared to the control. The extract treatment also brought about a non-significant increase in fructose content of the seminal vesicle and serum testosterone level. The findings of this study demonstrate that the extract acts in a dose-dependent manner, and it has varying effects on the reproductive parameters of male mice.

The current study established a HACCP tactic for all hazards related to Egyptian dry-cured pastrami production. All types of hazards that could occur at each production step were depicted. The fabrication steps of pastrami were originally based on the processes and conclusions presented in two previous publications by members of the research team; thus, the current scientific paper is considered a completion of the two previous publications. All operations executed and products manufactured outside the pastrami factory were excluded. The HACCP system was only applied to dry-cured pastrami production stages from receiving frozen raw meat and non-meat ingredients until packaging and storing the final product. Four CCPs were detected and taken into account. The permissible critical limits of additives and non-meat ingredients were considered. Suitable corrective actions were arranged. Regular HACCP plan review, proper recording of detected CCPs and critical limits were necessary for validation processes. Following up with the supply chain by obtaining the certified suppliers, together with the importance of the verification procedures of the elaborated HACCP plan, were essential in order to ensure the production of safe food without defects.

The number of reptiles owned as pets has risen worldwide. Additionally, urban expansion has resulted in more significant human encroachment and interactions with the habitats of captive reptiles. Between May and October 2022, 48 reptiles from pet shops and 69 from households were sampled in the Timișoara metropolitan area (western Romania). Three different sample types were collected from each reptile: oral cavity, skin, and cloacal swabs. Salmonella identification was based on ISO 6579-1:2017 (Annex D), a molecular testing method ( invA gene target), and strains were serotyped in accordance with the Kauffman-White-Le-Minor technique; the antibiotic susceptibility was assessed according to Decision 2013/652. This study showed that 43.28% of the pet reptiles examined from households and pet shops carried Salmonella spp. All of the strains isolated presented resistance to at least one antibiotic, and 79.32% (23/29) were multi-drug-resistant strains, with the most frequently observed resistances being to gentamicin, nitrofurantion, tobramycin, and trimethoprim-sulfamethoxazole. The findings of the study undertaken by our team reveal that reptile multi-drug-resistant Salmonella is present. Considering this aspect, the most effective way of preventing multi-drug-resistant Salmonella infections requires stringent hygiene control in reptile pet shops as well as ensuring proper animal handling once the animals leave the pet shop and are introduced into households.

Since 1983, when the first report of a human Cryptosporidium spp. infection was published in Romania, and until now, many studies on cryptosporidiosis have been published in our country, but most of them are in the Romanian language and in national journals less accessible to international scientific databases. Although the infection was first recognized as a problem in children or immunocompromised people or more of a problem in low-income or underdeveloped global countries, we have shown in this review that it can also occur in people with normal immunological function and that the epidemiology of our country can provide a theoretical basis for the formulation of a Cryptosporidium spp. prevention strategy. In addition, 9.1% of healthy children and 73% of immunocompromised children were observed to have Cryptosporidium spp. infections. Higher rates have also been reported in immunocompromised adults (1.8-50%). Analyzing the prevalence of Cryptosporidium spp. infection in animals, we found values of 28.52% in cattle, 18% in buffalo calves, between 27.8 and 60.4% in pigs, 52.7% in dogs, and 29.4% in cats. Furthermore, in Romania, the burden of cryptosporidiosis, including acute infections and long-term sequelae, is currently unknown.

Antibiotic resistance is rising to dangerously high levels throughout the world. To cope with this problem, scientists are working on CRISPR-based research so that antibiotic-resistant bacteria can be killed and attacked almost as quickly as antibiotic-sensitive bacteria. Nuclease activity is found in Cas9, which can be programmed with a specific target sequence. This mechanism will only attack pathogens in the microbiota while preserving commensal bacteria. This article portrays the delivery methods used in the CRISPR-Cas system, which are both viral and non-viral, along with its implications and challenges, such as microbial dysbiosis, off-target effects, and failure to counteract intracellular infections. CRISPR-based systems have a lot of applications, such as correcting mutations, developing diagnostics for infectious diseases, improving crops productions, improving breeding techniques, etc. In the future, CRISPR-based systems will revolutionize the world by curing diseases, improving agriculture, and repairing genetic disorders. Though all the drawbacks of the technology, CRISPR carries great potential; thus, the modification and consideration of some aspects could result in a mind-blowing technique to attain all the applications listed and present a game-changing potential.