Evaluation of Claw Lesions in Beef Cattle Slaughtered in Northern Portugal: A Preliminary Study.

Claw diseases have a profound impact on cattle welfare, affecting behaviors such as grazing, rumination, rest, decubitus, and water consumption. This study aimed to assess the prevalence of claw lesions and classify them according to the ICAR Claw Health Atlas (International Committee of Animal Recording) in two slaughterhouses. The influence of claw lesions on carcass weight, classification, and fat deposition was also examined. Involving 343 crossbreed cattle from 103 different extensive or semi-intensive farms, this study found an animal prevalence of claw disorders at 65.8%, with a higher incidence in females (n = 207, 60.35%) compared to males (n = 136, 39.65%). Despite the observed prevalence, claw lesions were not influenced by age or sex (p > 0.05). The main claw lesions identified, including heel horn erosion, double sole, and asymmetric claw, were consistent with the cattle management practices in the study area. These cattle were raised in small, rustic premises with uneven floors, utilizing a mix of manure and plant material as bedding and lacking access to pasture. Also, no negative economic impact was detected concerning carcass weight, classification, or fat deposition. Consequently, it was concluded that the presence of claw lesions in beef cattle raised under the characteristic management of this geographical area does not adversely affect animal health or farm economics.

Draft genome sequences of two Salmonella Uzaramo isolates from poultry in South Africa.

Salmonella enterica is a zoonotic pathogen and a leading cause of foodborne gastroenteritis in humans. Here, we report the draft genome sequences of two Salmonella Uzaramo isolates, which were isolated from poultry organs during routine post-mortem examination in South Africa. Currently, whole-genome sequences on Salmonella Uzaramo are scanty.

Antimicrobial growth promoters approved in food-producing animals in South Africa induce shiga toxin-converting bacteriophages from Escherichia coli O157:H7.

In this study, four antimicrobial growth promoters, including virginiamycin, josamycin, flavophospholipol, poly 2-propenal 2-propenoic acid and ultraviolet light, were tested for their capacity to induce stx-bacteriophages in 47 Shiga toxin-producing E. coli O157:H7 isolates. Induced bacteriophages were characterized for shiga toxin subtypes and structural genes by PCR, DNA restriction fragment length polymorphisms (RFLP) and morphological features by electron microscopy. Bacteriophages were induced from 72.3% (34/47) of the STEC O157:H7 isolates tested. Bacteriophage induction rates per induction method were as follows: ultraviolet light, 53.2% (25/47); poly 2-propenal 2-propenoic acid, 42.6% (20/47); virginiamycin, 34.0% (16/47); josamycin, 34.0% (16/47); and flavophospholipol, 29.8% (14/47). A total of 98 bacteriophages were isolated, but only 59 were digestible by NdeI, revealing 40 RFLP profiles which could be subdivided in 12 phylogenetic subgroups. Among the 98 bacteriophages, stx2a, stx2c and stx2d were present in 85.7%, 94.9% and 36.7% of bacteriophages, respectively. The Q, P, CIII, N1, N2 and IS1203 genes were found in 96.9%, 82.7%, 69.4%, 40.8%, 60.2% and 73.5% of the samples, respectively. Electron microscopy revealed four main representative morphologies which included three bacteriophages which all had long tails but different head morphologies: long hexagonal head, oval/oblong head and oval/circular head, and one bacteriophage with an icosahedral/hexagonal head with a short thick contractile tail. This study demonstrated that virginiamycin, josamycin, flavophospholipol and poly 2-propenal 2-propenoic acid induce genetically and morphologically diverse free stx-converting bacteriophages from STEC O157:H7. The possibility that these antimicrobial growth promoters may induce bacteriophages in vivo in animals and human hosts is a public health concern. Policies aimed at minimizing or banning the use of antimicrobial growth promoters should be promoted and implemented in countries where these compounds are still in use in animal agriculture.

Enterotoxigenic and Antimicrobic Susceptibility Profile of Staphylococcus aureus Isolates from Fresh Cheese in Croatia.

Certain Staphylococcus aureus strains harbour staphylococcal enterotoxin genes and hence can produce enterotoxin during their growth in food. Therefore, food can be a source of staphylococcal food poisoning, one of the most common food-borne diseases worldwide. Epidemiological data show that S. aureus is often present in raw milk cheeses, and consequently, cheeses are often the source of staphylococcal food poisoning outbreaks. The aim of this study was to determine the phenotypic characteristics of S. aureus isolates from fresh cheese, including antibiotic susceptibility; the presence of classical sea-see enterotoxin genes through molecular methods; and the isolate's ability to produce SEA-SEE enterotoxins in vitro through reversed passive latex agglutination. A total of 180 coagulase-positive staphylococci were isolated from 18 out of 30 cheese samples, and 175 were confirmed as S. aureus through latex agglutination and API STAPH tests. All isolates possessed phenotypic characteristics typical for S. aureus, with certain variations in the egg yolk reaction (18.3% of the isolates showed a weak reaction and 28% no reaction at all) and haemolysis pattern (36.6% of the isolates produced double-haemolysis and 4.6% were non-haemolytic). Antibiotic resistance was observed in 1.1% of the isolates and to mupirocin only. Real-time PCR detected the sec gene in 34 (19.4%) isolates, but most isolates (80.6%) were not enterotoxigenic. For all 34 (19.4%) strains that carried the sec gene, the RPLA method detected the production of the SEC enterotoxin in vitro. For those enterotoxigenic strains, the possibility of enterotoxin production in fresh cheese could not be ruled out.

Valorization of Pumpkin Byproducts: Antioxidant Activity and Carotenoid Characterization of Extracts from Peel and Filaments.

Pumpkin (Cucurbita sp.) represents an unquestionable source of valuable nutrients and bioactive compounds having a broad spectrum of health-promoting effects. The goal of this work was to characterize the byproducts (peels and filaments) of different pumpkin varieties belonging to C. moschata (Butternut, Lunga di Napoli, Moscata di Provenza, and Violina rugosa) and C. maxima (Delica, Delica vanity, Hokkaido, and Mantovana) species in terms of total carotenoid content, antioxidant activity, and carotenoid profiling. The research revealed that peels and filaments were a good source of ß-carotene and other non-esterified carotenoids, as well as esterified carotenoids. Considering the growing market demand for safe and healthy food products, pumpkin byproducts, having also an interesting antioxidant bioactivity, could be useful in the development of novel functional products.

The Importance of the Slaughterhouse in Surveilling Animal and Public Health: A Systematic Review.

From the point of public health, the objective of the slaughterhouse is to guarantee the safety of meat in which meat inspection represent an essential tool to control animal diseases and guarantee the public health. The slaughterhouse can be used as surveillance center for livestock diseases. However, other aspects related with animal and human health, such as epidemiology and disease control in primary production, control of animal welfare on the farm, surveillance of zoonotic agents responsible for food poisoning, as well as surveillance and control of antimicrobial resistance, can be monitored. These controls should not be seen as a last defensive barrier but rather as a complement to the controls carried out on the farm. Regarding the control of diseases in livestock, scientific research is scarce and outdated, not taking advantage of the potential for disease control. Animal welfare in primary production and during transport can be monitored throughout ante-mortem and post-mortem inspection at the slaughterhouse, providing valuable individual data on animal welfare. Surveillance and research regarding antimicrobial resistance (AMR) at slaughterhouses is scarce, mainly in cattle, sheep, and goats. However, most of the zoonotic pathogens are sensitive to the antibiotics studied. Moreover, the prevalence at the slaughterhouse of zoonotic and foodborne agents seems to be low, but a lack of harmonization in terms of control and communication may lead to underestimate its real prevalence.

Presence, Tissue Localization, and Gene Expression of the Adiponectin Receptor 1 in Testis and Accessory Glands of Male Rams during the Non-Breeding Season.

Adiponectin (ADIPOQ) is a member adipocytokines, and its actions are supported by two receptors, ADIPOQ receptor 1 and -2, respectively (ADIPOR1 and -R2). Our study was performed to evaluate the ADIPOR1 presence and location and its gene expression in reproductive tissues of the male ram, during its non-breading season. The different portions of the male ram reproductive system (testis, epididymis, seminal vesicle, ampoule vas deferens, bulb-urethral gland) were collected in a slaughterhouse. Immunohistochemistry showed ADIPOR1 positive signals in the cytoplasm of all the glandular epithelial cells, with a location near the nucleus; in the testes, the positive reaction was evidenced in the cytoplasm in the basal portion of the germinal epithelial cells. The immune reaction intensity was highest (p < 0.001) in the prostate and seminal vesicles glands than that of other parts of the ram reproductive tract. RT-qPCR detected the ADIPOR1 transcript in the testes, epididymis, vas deferens, bulbourethral glands, seminal vesicles, and prostate; the expression levels were high (p < 0.01) in the prostate and low (p < 0.01) in the testis, epididymis, and bulbourethral glands. The present results evidenced the possible ADIPOQ/ADIPOR1 system's role in regulating the testicular activity of male rams during the non-breading season.

Sex and seasonal differences in metal accumulation of selected tissues in red swamp crayfish from Lake Trasimeno (Umbria, Italy).

Concentrations of essential metals (Cu, Co, Cr, Fe, Ni, Mn, and Zn) and non-essential and toxic metals (Ag, Cd, Hg, and Pb) were quantified in hepatopancreas and abdominal muscle for 73 specimens (37 males and 36 females) of Procambarus clarkii from Lake Trasimeno (Italy) in a year's campaign (July 2018 to June 2019). Elemental concentrations determined in crayfish differed in tissues, seasons, and gender. Both sexes showed lower concentrations in abdominal muscle than in hepatopancreas. Essential metals accumulated in higher concentrations in both tissues than non-essential elements, while associations between essential metals were stronger, especially for females; however, a strong correlation was also observed between Cu and Ag in both sexes. No significant differences in concentrations were recorded between sexes for all seasons; contrarily, significant differences among the total seasonal concentrations (males plus females) were observed only for essential metals and for Ag. Seasonal Toxic Contamination Index (TCI) for Cr, Ni, Cu, Zn, Cd, Pb, and Hg were calculated to assess the toxicity level of metals in the sediments. TCI higher than 1 was detected for Ni only in summer, confirming the close connection between the fluctuating hydrological levels of the shallow Lake Trasimeno, strongly dependent on precipitation, and the metal contamination levels of sediments.

A quantitative risk assessment of Listeria monocytogenes from prevalence and concentration data: Application to a traditional ready to eat (RTE) meat product.

Ready to Eat (RTE) cooked meat products are among the most consumed RTE food subcategories in the EU/EEA. They are also associated with the highest number of identified listeriosis cases per year (>850), thus posing a public health risk especially among the susceptible population. This study estimated the risk of listeriosis from Italian head cheese (Coppa di Testa) consumption using a Quantitative Microbiological Risk Assessment (QMRA) based on data of prevalence and starting concentrations of Listeria monocytogenes in the product during a 3-year period (n = 1568). A consumer survey (n = 162) was conducted to provide information on domestic storage time and consumption habits, and storage conditions were determined from recordings of temperatures of domestic refrigerators (n = 57). A probabilistic model was designed for the evaluation of the growth of L. monocytogenes at each stage of the product pathway from production to consumption, using Monte Carlo simulations and employing the @Risk software. Risks associated to consumption of vacuum-packed and sliced-at-retail head cheese were assessed: The model predicted that the risk of listeriosis per serving of vacuum-packed product was in the 10-4 and 10-6 range (mean) for the high-risk and general populations respectively, and listeriosis cases were estimated to be greater than those due to consumption of sliced product (with risks in the range of 10-7 and 10-8). Overall, the model predicted that the mean number of listeriosis cases ranged from 0.001 to 0.24 and from 0.06 to 10 per one hundred thousand people, for the healthy and the high-risk population, respectively. Scenario analyses indicated that better control of the temperature of domestic refrigerators is effective in reducing the predicted risk of listeriosis for the longer stored vacuum-packed product by ~80 % for both the healthy and high-risk populations, whereas a shorter use-by-date of 30 days is an effective risk mitigation measure for both types of packed product. Model assumptions, as well as data gaps are discussed.

Apple Pomace as Valuable Food Ingredient for Enhancing Nutritional and Antioxidant Properties of Italian Salami.

Nowadays, food fortification with bioactive compounds deriving from agri-food waste is of great interest all over the world. In this work, apple pomace (AP), the most abundant by-product of apple juice manufacturing, was characterised by chemical, chromatographic and spectrophotometric analyses. AP showed valuable antioxidant activity, due to the presence of phenolic compounds (8.56 mg gallic acid equivalents/g), including quercetin-3-O-galactoside, quercetin-3-O-arabinofuranoside, and phloridzin. Dried AP, at 7% and 14%, was added to pork meat to produce Italian salami, then subjected to 25 days of ripening. Physicochemical, colorimetric and microbiological analyses were carried out at days 0, 5, 11, 19 and 25, while nutritional and sensory evaluations were performed at the end of the ripening. The overall acceptability was slightly higher for 7% AP compared to 14% AP sample, and generally the replacement of a percentage of meat with apple pomace allowed the production of salami with sensory properties comparable to those obtained with classic recipes. The improved fibre and phenol content, together with the lower fat and calories, represent the most interesting characteristics of fortified salami. The results confirm that the addition of AP represents a valid approach to adding healthy compounds to salami.

Effect of packaging and storage conditions on some quality traits of bovine meat.

Packaging is considered one of the most interesting technological aspects of food production and is a constantly evolving subject in food production. The type of packaging is important for the quality and safety of the product and for the visual appearance of the product to be immediately evaluated by consumers. The purpose of this study was to investigate the effect of four different types of modified atmosphere packaging (ATM) and vacuum packaging (VP) currently used by a company in central Italy, on the main qualitative characteristics of beef. For these two traditional and two new solutions with reduced environmental impact and compostable were evaluated. For each type of packaging, two different products were analyzed: steaks and hamburgers. The samples, immediately after production, were transported to the laboratory in refrigerated containers. Several parameters (color, pH, water holding capacity, drip loss, and microbiological characteristics) were evaluated at time 0 and after 7 (T7), 14 (T14) and 21 days (T21) of storage in the dark and at refrigeration temperature (+4°C ± 2°C). The results showed that the two types of packaging have very similar effects on the water-retaining capacity of the steaks. More noticeable differences were recorded by the colorimetric analysis: for both steaks and hamburgers, the products packaged in the traditional packaging appeared brighter and redder than those packaged in the new alternatives. The microbiological analysis of the steaks showed higher values in the "new" packaging. The formation of abundant ropy slime was observed in one of the samples in the "new" modified atmosphere package at T21. The results of this study showed that the technological characteristics (in particular, the color) and the microbiological characteristics of the steaks and hamburgers were better in "old" packaging, with a better appearance and a longer shelf life. The results obtained show how the research for eco-sustainable products for packaging must be addressed, taking into account the effect of the materials on the qualitative and hygienic- sanitary characteristics of the meat.

Delayed First Milking in Unassisted Overnight Calving Did Not Affect the Quality of Colostrum but Influenced Serum Brix Refractometry in Holstein Calves at Two Days of Life.

Timely administration of good-quality colostrum represents the first farm strategy to avoid the failure of passive transfer (FPT). However, calves born during the night are likely to be fed later than recommended. Our aim was to evaluate whether night-occurring calving and delayed first milking affected colostrum quality and immune passive transfer. The dataset included 463 calvings. Four liters of colostrum were administered by an esophageal tube feeder. The mean Brix% of colostrum was 27.43%, while serum Brix% at two days of life in calves was 10.19%. According to the Generalized Linear Model, parity ≥ 4, calving months of March, April, and from September to November positively influenced the quality of colostrum. Dams carrying a male calf produced lower quality colostrum compared with those carrying a female calf (−2.78 ± 1.04 Brix%, p = 0.008); heavier female calves were associated with greater colostrum quality (0.29 ± 0.05 for each kg increase, p < 0.001). Night- or day-calving had no effect on the quality of colostrum. The only factor influencing the serum Brix% of female Holstein calves at two days of life was the day- or night-occurring birth (−0.386 ± 0.188 Brix% in calves born during the night, p = 0.04). Our results showed that calves born overnight and fed the day after had decreased serum Total Protein concentrations as indicated by reduced Brix refractometer readings, compared with calves born during the day and fed quickly after birth. However, the administration of 4 L of high-quality colostrum likely improved their serum Brix% at two days of life. Alternatively, where the prevalence of good-quality colostrum is lower, improving calving supervision and ensuring timely feeding are important to reduce the risk of FPT.

Evolution and antimicrobial resistance of enterococci isolated from Pecorino and goat cheese manufactured on-farm in an area facing constraints as per EU Regulation 1305/2013 in Umbria, Italy.

The latest EU regulation on geographical indications (EU Regulation No. 1151/2012) has introduced a set of new tools for the protection and enhancement of food products in rural areas, under the group name of optional quality term (OQT). The Commission Delegated EU Regulation, No. 665/2014, regulated the conditions for the use of the optional quality term mountain product (MP), to support the implementation of a mountain value chain. This new tool is aimed at promoting local development, maintaining the economic activities in mountain areas, and redistributing wealth, whilst, at the same time, promoting the territory. Pecorino and goat cheeses are typical Italian cheeses made usually with whole raw ewe's or raw goat's milk, without starter culture addition. In an attempt to characterize these productions, the aim of this study was to investigate the evolution of enterococci during the production and ripening of Pecorino cheese made in three different farms, located in Umbria, Italy in areas facing natural or other specific constraints as stipulated by Regulation 1305/2013 on support for rural development by the European Agricultural Fund for Rural Development (EAFRD). Enterococci are enteric organisms which are commonly isolated from ewe and goat's milk production in Umbria, Italy. Counts of enterococci in raw milk ranged from 1.75 for ovine milk to 3.62 for ewe milk and a marked reduction was observed after thermization especially in ovine milk. Out of 100 isolates, 69 were E. faecium, 23 E. durans, 8 E. faecalis and 2 E. casseliflavus and the distribution of species between farms and between samples showed a prevalence of E. faecium in ovine farms and E. durans in ewes farms, with an equal dis-tribution between samples. High percentages of susceptible isolates were found for amoxicil-lin/clavulanic acid, ampicillin, chloramphenicol, sulphamethoxazole, sulphamethoxazole/ trimethoprim, ticarcillin, vancomycin. A high prevalence of resistant strains (>30%) was ob-served for amikacin, ciprofloxacin, ceftriaxone, kanamycin, tetracycline. A comparison of this re-sults with those of previous works on similar dairy products revealed high levels of resistance to antimicrobials which needs to be addressed.

Occurrence, Serotypes and Virulence Characteristics of Shiga-Toxin-Producing Escherichia coli Isolates from Goats on Communal Rangeland in South Africa.

Shiga-toxin-producing Escherichia coli is a foodborne pathogen commonly associated with human disease characterized by mild or bloody diarrhea hemorrhagic colitis and hemolytic uremic syndrome. This study investigated the occurrence of STEC in fecal samples of 289 goats in South Africa using microbiological culture and PCR. Furthermore, 628 goat STEC isolates were characterized by serotype (O:H) and major virulence factors by PCR. STEC was found in 80.2% (232/289) of goat fecal samples. Serotyping of 628 STEC isolates revealed 63 distinct serotypes including four of the major top seven STEC serogroups which were detected in 12.1% (35/289) of goats: O157:H7, 2.7% (8/289); O157:H8, 0.3%, (1/289); O157:H29, 0.3% (1/289); O103:H8, 7.6% (22/289); O103:H56, 0.3% (1/289); O26:H2, 0.3% (1/289); O111:H8, 0.3% (1/289) and 59 non-O157 STEC serotypes. Twenty-four of the sixty-three serotypes were previously associated with human disease. Virulence genes were distributed as follows: stx1, 60.6% (381/628); stx2, 72.7% (457/628); eaeA, 22.1% (139/628) and hlyA, 78.0% (490/628). Both stx1 and stx2 were found in 33.4% (210/628) of isolates. In conclusion, goats in South Africa are a reservoir and potential source of diverse STEC serotypes that are potentially virulent for humans. Further molecular characterization will be needed to fully assess the virulence potential of goat STEC isolates and their capacity to cause disease in humans.

A risk assessment model for Salmonella spp. in swine carcasses.

Salmonellosis is one of the most important food-borne outbreaks that occurs in the EU/EEA. From the first production stages at slaughter, meat is susceptible to spoilage and can be a substrate for the pathogenic microorganisms growth. Among the pathogens, the presence of Salmonella is mainly due to mishandling during the evisceration stage. For the year 2019, according to the collected data from MSs, on the 17.9% of all food-borne outbursts, the presence Salmonella was confirmed. Pork meat is considered as one of the four most commonly reported foods in cases of salmonellosis. For the training purposes of this project, Salmonella isolation and identification along with RA for carcass contamination, was performed. Pig carcasses were sampled using the non-destructive technique. The sampling took place post dressing and before the stage of chilling. For the Salmonella detection, a three phases process was performed (pre-enrichment, enrichment, isolation). A total of 757 samples were collected, 19 were found to be positive for Salmonella. The most common was found to be Salmonella Derby, which was identified eight times. The main objective of the project was to determine the prevalence of Salmonella spp. in swine carcasses. Moreover, certain parameters were evaluated in terms of their influence on the prevalence of Salmonella. A stochastic simulation model was developed in Microsoft Office Excel 2019 by using the add-in @Risk v.8.1. The prevalence was estimated to be 2.6%. For the pigs sampled, the average value of the distance from farm to slaughterhouse was 200.92 km. Additionally, the average weight of the carcasses was 127.97 kg. The prevalence of Salmonella between the samples that came from farms with a distance above the average, was higher by 1.7 units, while the prevalence for the samples with weight above the average was higher by 0.2 units. According to the stochastic model, it is specified that the prevalence is higher with greater distance, and there is an 8.1% probability the prevalence will exceed the legislation's - hygiene criteria. In addition, the prevalence of Salmonella was shown to increase, as well in the case of samples from weightier animals, but to a lesser extent.

Effect of Culling Management Practices on the Seroprevalence of Johne's Disease in Holstein Dairy Cattle in Central Italy.

A study was performed in Umbria, central Italy, to find out whether different culling strategies adopted by farms to control Johne's disease (JD) infection exerted effects on the seroprevalence in dairy cattle. Fifty Fresian dairy herds in the Perugia and Assisi districts were visited and an audit of herd management was conducted. Among the 50 herds, 20 were selected for the consistency of management practices and, according to the culling strategy, two groups were created: group A (aggressive culling protocol, with average herd productive life <1100 days) and group B (lower culling rate, with productive life greater than 1500 days). The presence of antibodies to Mycobacterium avium subspecies paratuberculosis (Map) in the serum was determined using a commercial enzyme-linked immunosorbent assay (ELISA) kit. It was found that 3.3% (n = 14) of the cows of group B (n = 422, from 17 herds) were positive for Map antibodies, in comparison with 5.7% (n = 21) of the cows from group A (n = 366, from three herds). The odds ratio from multiple logistic regression (adjusted odds ratio 2.446, 95% confidence interval 0.412 to 14.525) showed that Johne's disease prevalence in herds with a greater productive life was not higher than in herds with typical modern management characterized by more aggressive culling. This is a significant finding, indicating that aggressive culling may not be necessary. Current JD control recommendations are derived from data obtained in high-prevalence paratuberculosis areas (northern Europe, including northern Italy), while methods of information transfer to dairy farms in low-prevalence areas should be reassessed to ensure that the correct measures, including basic calving management and calf-rearing practices, are thoroughly implemented. Using the manufacturer's suggested cut-off for a positive ELISA test and the sensitivity and specificity claimed, the overall true prevalence in Umbria dairy cattle was calculated as 7% (95% confidence interval 5.2% to 8.8%).

Ultrasound-Assisted Extraction and Characterization of Polyphenols from Apple Pomace, Functional Ingredients for Beef Burger Fortification.

Currently, there is an increasing interest to valorise agri-food waste containing bioactive compounds with potential health benefits. In this paper, the recovery of functional molecules from apple pomace, the most abundant by-product of the apple processing industry, was carried out by ultrasound-assisted extraction (UAE) on fresh and freeze-dried samples. UAE extract, obtained by double extraction of freeze-dried apple pomace, was subjected to chromatographic and spectrophotometric characterization. It showed good levels of total phenol content, high antioxidant activity, and interesting antioxidant compounds (quercetin derivatives, chlorogenic acid, phloridzin). Subsequently, freeze-dried apple pomace, containing 40.19% of dietary fibre, was used as a fortifying agent for beef burgers (4% and 8%). The results concerning colour and sensory analysis of the fortified products were graded even better than the control (0%). The improved fibre and phenol content, together with the neutral flavour, represent the most interesting characteristics of fortified burgers. The results confirm that UAE was a successful technique for extracting phenol compounds and that the addition of apple pomace represents a valid approach to increase the health properties and palatability of beef burgers, including for consumers who do not like meat.

Estimating the Rates of Acquisition and loss of Resistance of Enterobacteriaceae to Antimicrobial Drugs in Pre-Weaned Dairy Calves.

The objective of this study was to investigate the effect of the antimicrobial drugs (AMD) on the shedding of resistant Enterobacteriaceae in feces of pre-weaned dairy calves. The AMD considered were ceftiofur, administered parenterally, and neomycin sulfate added in milk replacer and fed to calves during the first 20 days of life. Fifty-five calves, aged one to three days, were enrolled and followed to 64 days. Fecal samples were collected three times/week and treatments recorded daily. Enterobacteriaceae were quantified for a subset of 33 calves using spiral plating on plain, ceftiofur supplemented, and neomycin supplemented MacConkey agar. Negative binomial models were used to predict the association between treatment with AMD and the gain and loss of Enterobacteriaceae resistance over time. Acquisition of resistance by the Enterobacteriaceae occurred during treatment and peaked between days three to four post-treatment before decreasing to below treatment levels at days seven to eight post-treatment. Acquisition of neomycin resistance was observed on the first sampling day (day four from the start of feeding medicated milk replacer) to day eight, followed by cyclical peaks until day 29, when the Enterobacteriaceae counts decreased below pre-treatment. Enterobacteriaceae resistance against both AMD increased after AMD administration and didn't return to pre-therapeutic status until seven or more days after therapy had been discontinued. The study findings provide valuable insights into the dynamics of Enterobacteriaceae under routine AMD use in calves.

Antimicrobial Activity of Myrtus communis L. and Rosmarinus officinalis L. Essential Oils against Listeria monocytogenes in Cheese.

Listeria monocytogenes has been referred to as a concern microorganism in cheese making due to its ability to survive and grow in a wide range of environmental conditions, such as refrigeration temperatures, low pH and high salt concentration at the end of the production process. Since cheese may be a potential hazard for consumers, especially high-risk consumers (e.g., pregnant, young children, the elderly, people with medical conditions), efforts of the dairy industry have been aimed at investigating new conservation techniques based on natural additives to meet consumers' demands on less processed foods without compromising the food safety. Thus, the aim of this study was to evaluate the efficacy of Myrtus communis L. (myrtle) and Rosmarinus officinalis L. (rosemary) essential oils (EO) against Listeria monocytogenes ATCC 679 spiked in sheep cheese before ripening. After the cheesemaking process, the samples were stored at 8 °C for 2 h, 1 d, 3 d, 14 d and 28 d. The composition of EO was identified by gas chromatography-mass spectrometry (GC-MS) analysis. Constituents such as 1,8-cineole, limonene, methyl-eugenol, α-pinene, α-terpineol, α-terpinolene and ß-pinene were present in both EO, accounting for 44.61% and 39.76% from the total of chemical compounds identified for myrtle and rosemary EO, respectively. According to the chemical classification, both EO were mainly composed of monoterpenes. Minimum inhibitory concentration (MIC) against L. monocytogenes was obtained at 31.25 µL/mL to myrtle EO and at 0.40 µL/mL to rosemary EO. Then, cheeses were inoculated with L. monocytogenes (Ca. 6 log CFU/mL) and EO was added at MIC value. The addition of rosemary and myrtle EO displayed lower counts of L. monocytogenes (p < 0.01) (about 1-2 log CFU/g) during the ripening period compared to control samples. Ripening only influences (p < 0.001) the growth of L. monocytogenes in control samples. Since rosemary and myrtle EO do not exert any negative impact on the growth of native microflora (p > 0.05), their use as natural antimicrobial additives in cheese demonstrated a potential for dairy processors to assure safety against L. monocytogenes.

Effect of Delayed Refrigeration on the Microbial Carcass Contamination of Wild Boars (Sus scrofa).

The immediate refrigeration of meat after slaughter is a key issue for the proper storage and aging of meat. The industry standard cold chain relies on low temperatures and ventilation to lower the internal carcass temperature to 0-4 °C within the first 48 h, i.e., within four times the so-called semi-cooling time. On the other hand, for games, once bled and eviscerated, the carcass must be sent to a point where it can be sectioned or kept on air for maturation at refrigeration temperature. The precautions to observe are few and simple but essential: protect the meat and start the cooling process quickly. After preparing the animal (bleeding and evisceration), it may be necessary to face a period of transport that is sometimes long and not very easy; while small animals can be easily transported in a backpack, larger ones must necessarily be carried by several people or sometimes dragged to the vehicle capable of transporting them. It is obvious that a wild boar opened from the jaws to the pelvis and dragged for hundreds of meters will tend to be contaminated, although these contaminations are to be considered secondary for the preservation of the meat, compared to contamination by the intestinal contents. In an attempt to investigate the effect of delayed refrigeration on wild boar carcass contamination, the aim of this work was to determine a correlation between several hunting and logistic parameters (age, sex, animal weight, shooting distance, number of shots, weather and temperature and time from shot to refrigeration and to analysis) and bacterial contamination of the carcass. The correlation coefficient, r, was found to be 0.038 for the eviscerated body weight (p < 0.05), 0.091 for the external temperature on the day of hunting (p < 0.05), 0.027 for the time from shot to refrigeration (p = 0.081), 0.038 for the time from refrigeration to analysis (p < 0.05) and 0.043 for the time from shot to analysis (p < 0.05). These results stand for a negative correlation between the bacterial population and eviscerated carcass weight and between the bacterial population and external temperature and for a positive correlation between the time from shot to analysis and from refrigeration to analysis. No association was demonstrated between the bacterial population and the time from shot to refrigeration.