Effect of nickel on the comparative performance of inverse fluidized bed and continuously stirred tank reactors for biogenic sulphur-driven autotrophic denitrification.

The comparative performance of an inverse fluidized bed reactor (IFBR) having high density polyethylene beads as carrier materials for biofilm formation and a continuous stirred tank reactor (CSTR), both maintaining autotrophic denitrification using biogenic sulphur (ADBIOS) in the absence and presence of nickel (Ni2+), was studied. The reactors were compared in terms of NO3--N and NO2--N removal and SO42--S production throughout the study. A simulated wastewater with an inlet NO3--N concentration of 225 mg/L and a decreasing concentration of biogenic sulphur (bio-S) from 1.5 to 0.375 g/L was used. Both reactors were operated at a hydraulic retention time (HRT) of 48 h for 140 days and at an HRT of 42 h for the following 68 days. A more efficient ADBIOS was observed in the CSTR than IFBR throughout the study due to a better mixing of the feed wastewater in the bulk liquid and a higher availability of bio-S to the suspended cells. The NO3--N removal efficiency in the IFBR decreased by approximately 41% when the feed bio-S was reduced to 0.375 g/L, while it remained unaffected in the CSTR. Conversely, the presence of Ni2+ did not significantly affect NO3--N removal in both reactors even at a feed Ni2+ concentration of 120 mg/L. The highest NO3--N removal rates achieved were 86 and 108 mg NO3--N/(L·day) in the IFBR and CSTR, respectively, in the presence of 120 mg/L of feed Ni2+ at an HRT of 42 h. Batch studies conducted with acclimatized biomass showed that the continuous-flow operation mode in both reactors played a major role in helping the autotrophic denitrifiers to tolerate Ni2+ toxicity.

Detection of buffalo milk adulteration with cow milk by capillary electrophoresis analysis.

The addition of cow milk during the production of buffalo mozzarella is a common fraud in dairy industries because of the lower price and greater availability of cow milk throughout the year. The aim of this study was to develop a new, rapid, and robust capillary electrophoresis method for detecting and quantifying cow milk in buffalo milk by exploiting cow α-lactalbumin as a marker of adulteration. In particular, a linear calibration curve was generated, using a training set of calibrators consisting of 7 series of 17 buffalo/bovine whey mixtures, obtained after casein precipitation, with increasing percentages of cow whey. The capillary electrophoresis method showed high linearity (R2 = 0.968), repeatability [relative standard deviation (RSD) = 2.11, 3.02, 4.38, and 1.18%, respectively for 5, 10, 20, and 50% of buffalo/bovine whey mixtures], and intermediate precision (RSD = 2.18, 2.49, 5.09, and 3.19%, respectively, for 5, 10, 20, and 50% buffalo/bovine whey mixtures). Moreover, the minimum amount of detectable fraudulent cow milk was 1%, and the limit of quantification was 3.1%.

Application of high-intensity static magnetic field as a strategy to enhance the fertilizing potential of sewage sludge digestate.

Anaerobic digestion (AD) is a sustainable and well-established option to handle sewage sludge (SS), as it generates a methane-rich biogas and a digestate with potential fertilizing properties. In the past, different strategies have been proposed to enhance the valorization of SS. Among these, the application of a static magnetic field (SMF) has been poorly evaluated. This study aims to determine the effects of a high-intensity SMF (1.5 and 2 T) on the chemical composition of SS anaerobic digestate. Several strategies (i.e., number of magnetization cycles, addition of different sources and quantities of magnesium, and digestate aeration) have been applied to evaluate the possible formation of compounds with valuable fertilizing properties in the digestate. Experimental results showed that by combining different strategies promoting digestate exposure to the magnetic field it is possible to favour the reduction in the liquid phase of NH4+, NO3-, PO43-, SO42- and Mg2+ concentrations up to 28%, 38%, 34%, 39% and 31%, respectively. The XRD analyses conducted on the solid phase of the same magnetized digestate samples showed an increase in crystalline and amorphous phases of nitrogen and phosphorus compounds with fertilizing value, such as struvite. These results highlight that SMF application can increase the fertilizing potential of sewage sludge digestate and promote its valorization in a sustainable and circular perspective.

Evaluation of the Presence and Viability of Mycobacterium bovis in Wild Boar Meat and Meat-Based Preparations.

The aim of the present study is to provide information about the ability of Mycobacterium bovis to survive within wild boar (Sus scrofae) meat and meat-based preparations and the duration of this survival, and to consider the preservation of its infectious potential toward humans and animals. Meat samples were artificially contaminated with an M. bovis field strain and then stored at -20 °C, while two sausages batches were contaminated with the same field strain at two different concentrations, 105 CFU/g and 103 CFU/g, before storing them in proper conditions to allow for their ripening. A third sausage batch was contaminated by adding 2 g of wild boar lymph nodal tissue with active tuberculous lesions to the meat mixture. Bacteriological and biomolecular (PCR) methods were used to test the meat and sausage samples every 60 days and every 7-10 days, respectively. M. bovis was detected as still alive and viable on the frozen meat for the last test on the 342nd day, while from the sausage samples, M. bovis was isolated until 23 days after contamination. Our results indicate that M. bovis can stay alive and be viable for 23 days within sausages prepared with contaminated meat from infected wild boars. These products are usually eaten as fresh food after grilling, often cooking at a temperature that does not ensure complete inactivation of the pathogenic microorganisms present, which can pose a risk for humans to develop zoonotic tuberculosis.

Fungal Contamination and Aflatoxin B1 Detected in Hay for Dairy Cows in South Italy.

The growth of filamentous fungi on fodder is recognized as responsible for fungal deterioration and mycotoxin contamination of the plant mass leads to economic losses in the dairy cow production system. Mycotoxin contamination has significant implications for human and animal health and is one of the major concerns in the food and feed chain. This research provides an insight into the variety of viable molds (i.e., filamentous microfungi) that can be isolated from hay produced in South Italy and destined to dairy cows. On different lots of hay (n = 55) collected from 20 dairy farms, a total of 33 different fungal species were identified. The most representative was Cladosporium cladosporioides (n = 46, 84%) followed by Alternaria alternata (n = 25, 45%), and Rhizopus stolonifer (n = 24, 44%). The species most closely related to aflatoxin (AF) contamination, Aspergillus flavus, was often isolated (n = 11, 20%). Regarding AF detection, all the hay samples were found to be scarcely contaminated by AFB1 and showed values from 0.0020 to 0.0077 mg/kg, below the limits established by European Union (EU legislation) (0.02 mg/kg). None of the samples were positive for Aspergillia and tested for AFB1 showed results exceeding established limits. Additionally, hay with moisture between 15.0 and 19.2% or crude ash on dry matter content ranging from 14.0 to 15.5% reported an increased presence of AFB1 (p < 0.05) compared to the other samples. All the analyzed hay samples, besides the presence of molds, can be considered safe for the presence of AFB1. Prevention of mold spoilage is mandatory to reduce the exposure of humans and animals to mycotoxins.

S. aureus Biofilm Protein Expression Linked to Antimicrobial Resistance: A Proteomic Study.

Antimicrobial resistance (AMR) represents one of the most critical challenges that humanity will face in the following years. In this context, a "One Health" approach with an integrated multidisciplinary effort involving humans, animals and their surrounding environment is needed to tackle the spread of AMR. One of the most common ways for bacteria to live is to adhere to surfaces and form biofilms. Staphylococcus aureus (S. aureus) can form biofilm on most surfaces and in a wide heterogeneity of environmental conditions. The biofilm guarantees the survival of the S. aureus in harsh environmental conditions and represents an issue for the food industry and animal production. The identification and characterization of biofilm-related proteins may provide interesting insights into biofilm formation mechanisms in S. aureus. In this regard, the aims of this study were: (i) to use proteomics to compare proteomes of S. aureus growing in planktonic and biofilm forms in order to investigate the common features of biofilm formation properties of different strains; (ii) to identify specific biofilm mechanisms that may be involved in AMR. The proteomic analysis showed 14 differentially expressed proteins among biofilm and planktonic forms of S. aureus. Moreover, three proteins, such as alcohol dehydrogenase, ATP-dependent 6-phosphofructokinase, and fructose-bisphosphate aldolase, were only differentially expressed in strains classified as high biofilm producers. Differentially regulated catabolites metabolisms and the switch to lower oxygen-related metabolisms were related to the sessile conformation analyzed.

Foodborne Pathogen Assessment in Raw Milk Cheeses.

General hygienic parameters and selected foodborne pathogens in raw milk cheeses at the retail level were evaluated. A total of 245 raw milk cheese samples were analysed for total bacterial count, Enterobacteriaceae, E. coli, Salmonella spp., Listeria monocytogenes, coagulase-positive Staphylococci, and staphylococcal enterotoxin. Results showed only 3 samples that were not compliant with European rules on staphylococcal enterotoxin, but coagulase-positive Staphylococci were evidenced in all samples tested. Salmonella spp. and Listeria monocytogenes were never detected whereas E. coli was evidenced in 20 samples. Results suggest a need for improvement of good manufacturing practice and milking operation.

Rapid Liquid AP-MALDI MS Profiling of Lipids and Proteins from Goat and Sheep Milk for Speciation and Colostrum Analysis.

Rapid profiling of the biomolecular components of milk can be useful for food quality assessment and for food fraud detection. Differences in commercial value and availability of milk from specific species are often the reasons for the illicit and fraudulent sale of milk whose species origin is wrongly declared. In this study, a fast, MS-based speciation method is presented to distinguish sheep from goat milk and sheep colostrum at different phases. Using liquid atmospheric pressure (AP)-matrix-assisted laser desorption/ionisation (MALDI) MS, it was possible to classify samples of goat and sheep milk with 100% accuracy in one minute of data acquisition per sample. Moreover, an accuracy of 98% was achieved in classifying pure sheep milk samples and sheep milk samples containing 10% goat milk. Evaluating colostrum quality and postnatal stages represents another possible application of this technology. Classification of sheep colostrum samples that were collected within 6 hours after parturition and 48 hours later was achieved with an accuracy of 84.4%. Our data show that substantial changes in the lipid profile can account for the accurate classification of colostrum collected at the early and late time points. This method applied to the analysis of protein orthologs of different species can, as in this case, allow unequivocal speciation analysis.

Milk microbiota: Characterization methods and role in cheese production.

Milk is a complex body fluid aimed at addressing the nutritional and defensive needs of the mammal's newborns. Harbored microbiota plays a pivotal role throughout the cheesemaking process and contributes to the development of flavor and texture typical of different type of cheeses. Understanding the dairy microbiota dynamics is of paramount importance for controlling the qualitative, sensorial and biosafety features of the dairy products. Although many studies investigated the contribution of single or few microorganisms, still there is some information lacking about microbial communities. The widespread of the omics platforms and bioinformatic tools enable the investigation of the cheese-associated microbial community in both phylogenetical and functional terms, highlighting the effects of the diverse cheesemaking variables. In this review, the most relevant literature is revised to provide an introduction of the milk- and cheese-associated microbiota, along with their structural and functional dynamics in relation to the diverse cheesemaking technologies and influencing variables. Also, we focus our attention on the latest omics technologies adopted in dairy microbiota investigation. Discussion on the key-steps and major drawbacks of each omics discipline is provided along with a collection of results from the latest research studies performed to unravel the fascinating world of the dairy-associated microbiota. SIGNIFICANCE: Understanding the milk- and cheese- associated microbial community is nowadays considered a key factor in the dairy industry, since it allows a comprehensive knowledge on how all phases of the cheesemaking process impact the harbored microflora; thus, predict the consequences in the finished products in terms of texture, organoleptic characteristics, palatability and biosafety. This review, collect the pioneering and milestones works so far performed in the field of dairy microbiota, and provide the basic guidance to whom approaching the cheese microbiota investigation by means of the latest omics technologies. Also, the review emphasizes the benefits and drawbacks of the omics disciplines, and underline how the integration of diverse omics sciences enhance a comprehensive depiction of the cheese microbiota. In turn, a better consciousness of the dairy microbiota might results in the application of improved starter cultures, cheesemaking practices and technologies; supporting a bio-safe and standardized production of cheese, with a strong economic benefit for both large-scale industries and local traditional dairy farms.

Evaluation of freezing point in milk from buffalos reared in Calabria, Italy.

Evaluation of freezing point is one of the most common technique used to detect milk adulteration such as addition of external water to increase volume. The aim of this study was to evaluate the freezing point of buffalo milk using infrared spectroscopy and to assess how it is influenced by other milk components. A total of 361 individual buffalo milk samples were collected monthly from March to August of 2017 in a dairy farm in Catanzaro district, Italy. Samples were tested for freezing point, urea, acetone and beta-hydroxybutyrate, percent of fat, protein, lactose, casein, by Fourier Transformed Spectroscopy. The pH and daily milk production were also recorded. Freezing point ranged from -0.574°C to - 0.512°C and the mean values was -0.545°C ±0.010. According to lactation stage, freezing point decreased until 210 days post-partum reaching the minimum value of -0.550°C, then it slightly increased during lactation; according to sampling month the highest and lowest values were recorded in August and June, respectively. A positive correlation between freezing point and lactose content were evidenced (r=0.1806, P<0.05). Moreover, a faintly positive correlation was also found between freezing point and beta-idroxibutirrate (r=0.0869, P<0.05) and acetone (r=0.0096, P<0.05), whereas a negative correlation with fat (r=-0.2356, P<0.05), protein (r=-0.1855, P<0.05), casein (r=-0.2127, P<0.05) and urea (r=-0.1229, P<0.05) was evidenced.

Phenotypic Antimicrobial Resistance Profile of Isolates Causing Clinical Mastitis in Dairy Animals.

Mastitis is the most frequent and costly disease of lactating animals and is associated with a significant reduction in milk yield, increased cost and culling. Early and specific antibiotic based treatment reduces the severity of the disease. Over the years the extensive use of antimicrobials has led to increase antimicrobial resistance. The present study was designed to investigate the prevalence of microorganisms responsible for mastitis and their antimicrobial resistance pattern. A total of 282 milk samples were collected from different animal species (sheep, cows and goats) with clinical mastitis. Antimicrobial resistance was evaluated for Streptococcus spp. and Staphylococcus spp. In cow samples Streptococcus spp. represented the most frequently isolated genus (33.84%), while Staphylococcus spp. was the most prevalent genus in sheep and goat samples (44.4 and 73.86%, respectively). Gentamicin and chloramphenicol were found to be the most effective drugs against the tested isolates, while the highest resistance rates were observed for amoxicillin, ampicillin, tetracycline, trimethoprim-sulfamethoxazole.

Cystic echinococcosis in cattle dairy farms: spatial distribution and epidemiological dynamics.

A study monitoring cystic echinococcosis (CE) in adult dairy cattle from intensive livestock farms located in the municipality of Arborea (Sardinia, Italy) was carried out between 2012 and 2015. A retrospective study of veterinary reporting forms of post-mortem inspections in 10 different Italian slaughterhouses was also performed. In addition, data on viability and molecular characterisation of hydatid cysts removed from parasitised organs in cattle was carried out. A geographical information system (GIS) with data layers of the study area and the geo-referenced points of 160 cattle farms was constructed. CE was found in 21.9% (35/160) of the surveyed farms. The retrospective study revealed that 0.05% (13/23,656) of adult slaughtered animals (over one year of age) from Arborea had tested positive to CE. The results stratified per year showed the following CE prevalences: 0.09% (5/5673) in 2012; 0.02% (1/5682) in 2013; 0.08% (5/6261) in 2014; and 0.03% (2/6040) in 2015 (χ2 with 3 degrees of freedom=3.81; P=0.282). The E. granulosus sensu stricto (formerly called G1 or sheep strain) was detected in all cysts subjected to molecular analysis. The GIS analysis showed that CE is fairly resilient in the Arborea territory where most of cattle farms are located, while a small cluster of cases was found located in the southeastern part of Arborea, close to districts where sheep farms are situated. The present survey reports the presence of CE in Sardinian dairy cattle intensive farms and suggests that the parasitic pressure of CE in the island continues to be very strong.

Serological and Molecular Investigation of Swine Hepatitis E Virus in Pigs Raised in Southern Italy.

Hepatitis E virus (HEV) infection is a common acute hepatitis transmitted by the fecal-oral route. In developed countries, the virus has a zoonotic potential, and domestic pigs and wild boars are considered main reservoirs. To assess the prevalence of HEV-positive animals in the Calabria region (southern Italy) on a serological and molecular level, a total of 216 autochthonous healthy pigs (Apulo-Calabrese breed) were sampled. Both sera and feces were collected. Pigs were grouped based on age: 117 pigs <6 months and 99 pigs >6 months. By using a commercial enzyme-linked immunosorbent assay system, a total of 173 (80%) of the 216 pigs tested seropositive. In all sampled farms (n = 8), pigs with antibodies (immunoglobulin G) against HEV were detected at a level higher than 60%, with a significant difference among age groups (P < 0.0001). Moreover, 16 fattening pigs were found to be nested reverse transcription PCR positive and thus to shed viral genomes in their feces. These positive findings resulted in a prevalence of 48.4% on the farm level (16 of 35 pigs) and an overall prevalence of 7.4% at the animal level (16 of 216 pigs). Based on the present study, HEV seems to circulate among the autochthonous domestic pig population of southern Italy with a low sharing rate. Further studies exploring the origin of infection are needed to minimize the risk of human exposure and to reduce consequences for public health.

Use of Sonoelastography to Evaluate Texture Modifications of Mozzarella di Bufala Campana Protected Designation of Origin During Storage at Different Temperatures.

Mozzarella cheese from buffalo milk is a fresh, stringy-textured dairy product, exhibiting a porcelain white colour, a smooth, bright, and humid surface, an extremely thin rind and delicate taste. The high humidity typical of this cheese reduces its shelf-life and it is cause of dramatic organoleptic changes during storage. In this study we tested sonoelastography to evaluate texture changes of mozzarella cheese from buffalo milk during storage. Cheeses form local market produced in the same condition were divided in three batches and stored in different conditions: the first (B1) was stored in preserving liquid at room temperature (20°C); the second (B2) was stored without preserving liquid at 4°C; and the third (B3) was stored at 4°C in preserving liquid. In B1 sonoelastography showed a reduction of the hardness and stiffness of rind, while in B2 inelastic tissue increased its thickness. Best results were obtained in B3, where no significant difference was evidenced during storage.