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.

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 Regarding the control of diseases in livestock, scientific research is scarce and outdated, not taking advantage of the potential for disease control.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.

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.

Antimicrobial Resistance and Clonal Lineages of Staphylococcus aureus from Cattle, Their Handlers, and Their Surroundings: A Cross-Sectional Study from the One Health Perspective.

Staphylococcus aureus have been progressively identified in farm animals and in humans with direct contact with these animals showing that S. aureus may be a major zoonotic pathogen. Therefore, we aimed to isolate S. aureus from cows, their handlers, and their immediate surroundings, and to investigate the antimicrobial resistance and genetic lineages of the isolates. Mouth and nose swabs of 244 healthy cows (195 Maronesa, 11 Holstein-Friesians, and 28 crossbreeds), 82 farm workers, 53 water and 63 soil samples were collected. Identification of species was carried out by MALDI-TOF MS Biotyper. The presence of antimicrobial resistance genes and virulence factors was assessed based on gene search by PCR. All isolates were typed by multilocus sequence typing and spa-typing. From 442 samples, 33 (13.9%), 24 (29.3%), 1 (2%), and 1 (2%) S. aureus were recovered from cows, farm workers, water, and soil samples, respectively. Most of the isolates showed resistance only to penicillin. S. aureus isolates were ascribed to 17 sequence types (STs) and 26 spa-types. Some clonal lineages were common to both cows and farm workers such as ST30-t9413, ST72-t148, and ST45-t350. Through a One Health approach, this study revealed that there is a great diversity of clonal lineages of S. aureus in cows and their handlers. Furthermore, some S. aureus lineages are common to cows and handlers, which may suggest a possible transmission.

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.

Use of Starter Cultures in Foods from Animal Origin to Improve Their Safety.

Starter cultures can be defined as preparations with a large number of cells that include a single type or a mixture of two or more microorganisms that are added to foods in order to take advantage of the compounds or products derived from their metabolism or enzymatic activity. In foods from animal origin, starter cultures are widely used in the dairy industry for cheese, yogurt and other fermented dairy products, in the meat industry, mainly for sausage manufacture, and in the fishery industry for fermented fish products. Usually, microorganisms selected as starter culture are isolated from the native microbiota of traditional products since they are well adapted to the environmental conditions of food processing and are responsible to confer specific appearance, texture, aroma and flavour characteristics. The main function of starter cultures used in food from animal origin, mainly represented by lactic acid bacteria, consists in the rapid production of lactic acid, which causes a reduction in pH, inhibiting the growth of pathogenic and spoilage microorganisms, increasing the shelf-life of fermented foods. Also, production of other metabolites (e.g., lactic acid, acetic acid, propionic acid, benzoic acid, hydrogen peroxide or bacteriocins) improves the safety of foods. Since starter cultures have become the predominant microbiota, it allows food processors to control the fermentation processes, excluding the undesirable flora and decreasing hygienic and manufacturing risks due to deficiencies of microbial origin. Also, stater cultures play an important role in the chemical safety of fermented foods by reduction of biogenic amine and polycyclic aromatic hydrocarbons contents. The present review discusses how starter cultures contribute to improve the microbiological and chemical safety in products of animal origin, namely meat, dairy and fishery products.

Consumer Knowledge about Food Labeling and Fraud.

Food fraud is a growing problem and happens in many ways including mislabelling. Since lack of consumers' knowledge about mandatory food labeling information and different types of food fraud may impact public health, the present work assesses consumers' knowledge about these issues. Principal component analysis was performed to obtain a smaller number of uncorrelated factors regarding the usefulness and confidence of information displayed in food labels and the perception of food fraud. Results indicated that information displayed in food labels is useful, however the way it is presented may decrease consumer interest and understanding. Regarding respondents' confidence in foodstuffs, over half of them stated that information provided in food labels is reliable. However, a lack of confidence about food composition is observed in those processed foodstuffs such as meat products. Food fraud is recognized by more than half of respondents with a higher perception of those practices that imply a risk to public health than those related to economic motivation. Age and education of consumers influenced the perception of the information displayed in the food labels, their confidence and knowledge about food fraud. Implementation of education programs to increase consumer knowledge about food labelling and fraud is essential. Respondents' perception results could be use as guidelines by the food industry to improve food label design in order to enhance consumer understanding.

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.

Microbiological Quality of Foodstuffs Sold on Expiry Date at Retail in Portugal: A Preliminary Study.

Currently, food waste represents an important issue due to its negative economic, social and environmental impact. To reduce the food waste levels, some retailers' brands implement discounting based on the proximity to expiry. Since this practice may involve potential food poisoning, a total of 94 food products from animal origin, purchased in two supermarkets in North Portugal on the expiry date, were analyzed for selected foodborne and spoilage microorganisms. Moreover, the samples were classified as satisfactory and not satisfactory according to their microbiological quality. The results showed that none of the samples presented counts for Salmonella spp., S. aureus, B. cereus. L. monocytogenes was detected in one sample over the limit of 2 log cfu/g as defined by Regulation 2073/2005. The evaluation of food hygiene and spoilage indicators showed that the processed foods displayed lower counts than raw products (beef, pork, chicken and fish). Regarding Enterobacteriaceae, raw products presented on average over 2 log cfu/g than processed foods, with the exception of beef samples that accounted over 3 log cfu/g more than processed foods. In addition, E. coli was mainly detected in fresh meat of which chicken and pork displayed the highest counts. Regarding the qualitative classification, 51.06% of the samples were not satisfactory for the total mesophilic counts, while 62.76% and 58.51% displayed positive results for Enterobacteriaceae and molds and yeasts (M&Y) criteria, respectively. In all, 70.21% of the samples analyzed at the expiry date failed, at least, in one microbiological criterion. The results indicate that the foods available at the end of the shelf life in supermarkets do not represent a risk for food poisoning due to the absence of foodborne pathogens. Since the microbiological indicators of storage/handling of raw products were mainly unsatisfactory, this indicates that the sale of these perishable foods at the end of the shelf life may not be recommended. On the other hand, processed products subjected to food conservation procedures (i.e., thermal processing) could be sold at the end of their shelf life or donated beyond the best-before date, due to its physical, chemical and microbiological stability. However, evidences of foodborne outbreaks associated to this kind of foodstuffs indicated the need of a proper risk assessment. Moreover, it is important to remark that other factors such as small sample size, the absence of the evaluation of the handling, and storage conditions along the food chain or organoleptic alterations must be assessed in further studies.

Farm practices and risk factors which influence the high prevalence of brucellosis in small ruminant flocks in Northeast Portugal.

The region of Tras-os-Montes e Alto Douro in northeast Portugal displayed the highest prevalence of brucellosis in small ruminants of the country. A vaccination programme of the whole population with Brucella melitensis Rev-1 was carried out from 2001 to 2004 in an attempt to reduce prevalence levels and protect public health. Although individual prevalence decreased from 5.6% in 2001 to 0.4% in 2007, several flocks continued to present individual prevalence ≥ 5.0%. Given the multifactorial characteristics of brucellosis, the current study evaluated farming practices and risk factors in flocks with an individual prevalence over 5% by multifactorial correspondence analysis. Results showed that a lack of recognition of the symptoms of brucellosis and lack of Rev-1 vaccination were the main factors contributing to the high individual prevalence of brucellosis in flocks. Other factors such as the consumption of raw milk, presence of dog commingling with animals and use of communal pastures also contributed to the persistence of the disease. Family farms with low economical profit, minimal training/education of farmers, and a scarcity of veterinary support may explain the persistence of factors contributing to the high prevalence of brucellosis. The results of this study highlight several risk factors and farming practices that might have contributed to the maintenance of a high prevalence of brucellosis in flocks with high brucellosis prevalence. These results could be used to adopt new approaches to improve the efficiency of brucellosis eradication programs.

One Health Approach Reveals the Absence of Methicillin-Resistant Staphylococcus aureus in Autochthonous Cattle and Their Environments.

Antimicrobial resistance represents one of the greatest challenges of the twenty-first century, and it is globally recognized that addressing this problem requires a concerted One Health approach involving humans, animals, and the environment. Methicillin-resistant Staphylococcus aureus (MRSA) currently represents a global burden; it is resistant to almost all beta-lactams and some MRSA strains are highly multiresistant. S. aureus infection in cattle results in major economic losses in the food industry. Moreover, cases of livestock-associated MRSA strains responsible for invasive life-threatening infections have been reported among human patients in contact with infected or colonized animals. The autochthonous Maronesa cattle breed is a threatened rustic traditional Portuguese breed of mountain cattle of high importance for the Vila Real region. It has been used for centuries as motive power in all kinds of agricultural work and also for meat production, which is its current dominant use and the main source of economic value, being the Maronesa meat commercialized with PDO - Protected Designation of Origin. This study aimed to determine the prevalence and transmission of MRSA in cattle of the Maronesa breed, through a concerted One Health approach comprising human, water, and soil samples of the animals' handlers and environments. In a total of 195, 63, 40, and 43 cattle, human, water, and soil samples screened in selective ORSAB media supplemented with 2 mg/L oxacillin; only one human sample harbored a MRSA isolate which was ascribed to spa-type t9413 and to ST30, one of the most common genetic lineages associated with community-acquired MRSA. Considering the increasing reports of MRSA isolation from cattle and handlers in Europe, the absence of this major human and animal pathogen in Maronesa cattle and their production systems represents a serendipitous result, valuing this important autochthonous breed. To our knowledge, this is the first study to determine MRSA prevalence and transmission in Maronesa cattle. Through a concerted One Health approach, this study revealed that the Maronesa cattle and their surrounding environments do not represent reservoirs for Methicillin-resistant Staphylococcus aureus.

Synergistic Activity of Essential Oils from Herbs and Spices Used on Meat Products against Food Borne Pathogens.

Essential oils (EOs) could be utilized as natural agents to improve the safety of meat products. However, the high concentration required to achieve an antimicrobial effect in foods might be incompatible with their sensory acceptance. To avoid this problem, combinations of EOs provide an effective approach reducing the odds of sensory rejection. In our study, 13 EOs of herbs and spices commonly used in the seasoning of meat products were assessed for their antimicrobial activity against Salmonella spp., Listeria monocytogenes, Escherichia coli and Staphylococcus aureus. However, only 7 of them were selected to study their synergistic effect based on their antimicrobial activity and minimum inhibitory concentration (MIC) against foodborne pathogens. EOs of thyme and cinnamon presented the largest antibacterial activity against foodborne pathogens. Combinations of selected EOs displayed a synergic effect against foodborne pathogens and also an important decrease in their individual MIC. Thyme EO presented the lowest individual MIC, but its utilization in combination decreased the MIC of the other EOs. Utilization of cinnamon EO also improved the reduction of the individual MICs of the EOs of cumin and parsley. Our results suggest the potential use of EO mixtures to control foodborne pathogens in meat products. Although the individual MIC values of selected EOs decreased, the sensory impact on meat products needs to be assessed.

Influence of Food Characteristics and Food Additives on the Antimicrobial Effect of Garlic and Oregano Essential Oils.

Utilization of essential oils (EOs) as antimicrobial agents against foodborne disease has gained importance, for their use as natural preservatives. Since potential interactions between EOs and food characteristics may affect their antimicrobial properties, the present work studies the influence of fat, protein, pH, aw and food additives on the antimicrobial effect of oregano and garlic EOs against Salmonella spp. and Listeria monocytogenes. Results showed that protein, pH, aw, presence of beef extract, sodium lactate and nitrates did not influence their antimicrobial effect. In contrast, the presence of pork fat had a negative effect against both EOs associated with their dilution of the lipid content. The addition of food phosphates also exerts a negative effect against EOs probably associated with their emulsification properties as observed with the addition of fat. The results may help the food industry to select more appropriate challenges to guarantee the food safety of foodstuffs.

Biological Risks and Laboratory-Acquired Infections: A Reality That Cannot be Ignored in Health Biotechnology.

Advances and research in biotechnology have applications over a wide range of areas, such as microbiology, medicine, the food industry, agriculture, genetically modified organisms, and nanotechnology, among others. However, research with pathogenic agents, such as virus, parasites, fungi, rickettsia, bacterial microorganisms, or genetic modified organisms, has generated concern because of their potential biological risk - not only for people, but also for the environment due to their unpredictable behavior. In addition, concern for biosafety is associated with the emergence of new diseases or re-emergence of diseases that were already under control. Biotechnology laboratories require biosafety measures designed to protect their staff, the population, and the environment, which may be exposed to hazardous organisms and materials. Laboratory staff training and education is essential, not only to acquire a good understanding about the direct handling of hazardous biological agents but also knowledge of the epidemiology, pathogenicity, and human susceptibility to the biological materials used in research. Biological risk can be reduced and controlled by the correct application of internationally recognized procedures such as proper microbiological techniques, proper containment apparatus, adequate facilities, protective barriers, and special training and education of laboratory workers. To avoid occupational infections, knowledge about standardized microbiological procedures and techniques and the use of containment devices, facilities, and protective barriers is necessary. Training and education about the epidemiology, pathogenicity, and biohazards of the microorganisms involved may prevent or decrease the risk. In this way, the scientific community may benefit from the lessons learned in the past to anticipate future problems.