Addition to Thermized Milk of Lactococcus lactis subsp. cremoris M104, a Wild, Novel Nisin A–Producing Strain, Replaces the Natural Antilisterial Activity of the Autochthonous Raw Milk Microbiota Reduced by Thermization

2014 ◽  
Vol 77 (8) ◽  
pp. 1289-1297 ◽  
Author(s):  
ALEXANDRA LIANOU ◽  
JOHN SAMELIS

Recent research has shown that mild milk thermization treatments routinely used in traditional Greek cheese production are efficient to inactivate Listeria monocytogenes and other pathogenic or undesirable bacteria, but they also inactivate a great part of the autochthonous antagonistic microbiota of raw milk. Therefore, in this study, the antilisterial activity of raw or thermized (63°C, 30 s) milk in the presence or absence of Lactococcus lactis subsp. cremoris M104, a wild, novel, nisin A–producing (Nis-A+) raw milk isolate, was assessed. Bulk milk samples were taken from a local cheese plant before or after thermization and were inoculated with a five-strain cocktail of L. monocytogenes (approximately 4 log CFU/ml) or with the cocktail, as above, plus the Nis-A+ strain (approximately 6 log CFU/ml) as a bioprotective culture. Heat-sterilized (121°C, 5 min) raw milk inoculated with L. monocytogenes was used as a control treatment. All milk samples were incubated at 37°C for 6 h and then at 18°C for an additional 66 h. L. monocytogenes grew abundantly (>8 log CFU/ml) in heat-sterilized milk, whereas its growth was completely inhibited in all raw milk samples. Conversely, in thermized milk, L. monocytogenes increased by 2 log CFU/ml in the absence of strain M104, whereas its growth was completely inhibited in the presence of strain M104. Furthermore, nisin activity was detected only in milk samples inoculated with strain M104. Thus, postthermal supplementation of thermized bulk milk with bioprotective L. lactis subsp. cremoris cultures replaces the natural antilisterial activity of raw milk reduced by thermization.

2017 ◽  
Vol 41 (1) ◽  
pp. 49-54
Author(s):  
Sara Ahmed Abd Al-Zahra

      A total of 163 milk samples (500 ml each) of cows, ewe, goats, buffaloes, camels were collected randomly at weekly intervals (10 samples/ week) from both Abu-Ghraib and Al-Fudhailiya villages and also for different local retail markets inside the Baghdad province and these samples were including milk of different animal species, milk cans, bulk milk tanks and imported Ultra-high temperature processing milk. Among the total milk samples only 138 milk samples were examined during two climatic periods where the first period was in summer that extended from the beginning of September to the end of October 2015 while the second period was in winter that extended from the beginning of January to the end of February 2016. Besides that, some of the selected positive samples for Deltametrin residues were subjected to one of the commercial heat treatments such as 63°C/30 min., 80°C/5 min. and 100°C/5 min. to evaluate the efficiency of heat exposure on the degradation of Deltametrin residues in milk. The results pointed out that milk samples containing the higher fat percentage exhibited significantly (P<0.05) the highest concentration of Deltametrin in summer (0.08ppm) than in winter (0.008 ppm) seasons. It was clearly obvious that the detectable concentrations of the Deltametrin were higher in buffalo's and ewe milk samples than those found in cows, goats and camels and such results could be attributed to the higher fat content of buffalos and ewes milk than the other animals as well as the lipophilic nature of the Deltamethrin. In other word, increased the fat percentages of milk was being associated with an increased level of Deltametrin residues due to the lipophilic nature of the Deltametrin pesticide. The current results revealed that milk samples that were collected from buffaloes, ewes and cows recorded significantly (P<0.05) the highest Deltametrin residues in summer season where their mean levels that exceeded the accepted MRLs of 0.05 ppm to milk samples of goats and camels that had significantly (P<0.05) the lowest mean levels of Deltametrin residues where their means levels were 0.038 and 0.032 ppm respectively. There was a significant (P<0.05) seasonal variation of the Deltametrin concentrations in milk samples for each animal species where all the milk samples that were collected from buffaloes, ewe, cows, goats and camels had significantly (P<0.05) higher mean levels of Deltametrin residues in summer season than in winter season, milk samples that were collected from milk cans (5, 25 and 50 kg) recorded significantly (P<0.05). The highest Deltametrin residues during the summer season in comparison to 10 tons bulk milk tank samples.


2017 ◽  
Vol 80 (12) ◽  
pp. 2137-2146 ◽  
Author(s):  
Dimitrios Noutsopoulos ◽  
Athanasia Kakouri ◽  
Eleftheria Kartezini ◽  
Dimitrios Pappas ◽  
Efstathios Hatziloukas ◽  
...  

ABSTRACT This study evaluated in situ expression of the nisA gene by an indigenous, nisin A–producing (NisA+) Lactococcus lactis subsp. cremoris raw milk genotype, represented by strain M78, in traditional Greek Graviera cheeses under real factory-scale manufacturing and ripening conditions. Cheeses were produced with added a mixed thermophilic and mesophilic commercial starter culture (CSC) or with the CSC plus strain M78 (CSC+M78). Cheeses were sampled after curd cooking (day 0), fermentation of the unsalted molds for 24 h (day 1), brining (day 7), and ripening of the brined molds (14 to 15 kg each) for 30 days in a fully controlled industrial room (16.5°C; 91% relative humidity; day 37). Total RNA was directly extracted from the cheese samples, and the expression of nisA gene was evaluated by real-time reverse transcription PCR (qRT-PCR). Agar overlay and well diffusion bioassays were correspondingly used for in situ detection of the M78 NisA+ colonies in the cheese agar plates and antilisterial activity in whole-cheese slurry samples, respectively. Agar overlay assays showed good growth (&gt;8 log CFU/g of cheese) of the NisA+ strain M78 in coculture with the CSC and vice versa. The nisA expression was detected in CSC+M78 cheese samples only, with its expression levels being the highest (16-fold increase compared with those of the control gene) on day 1, followed by significant reduction on day 7 and almost negligible expression on day 37. Based on the results, certain intrinsic and mainly implicit hurdle factors appeared to reduce growth prevalence rates and decrease nisA gene expression, as well as the nisin A–mediated antilisterial activities of the NisA+ strain M78 postfermentation. To our knowledge, this is the first report on quantitative expression of the nisA gene in a Greek cooked hard cheese during commercial manufacturing and ripening conditions by using a novel, rarely isolated, indigenous NisA+ L. lactis subsp. cremoris genotype as costarter culture.


1996 ◽  
Vol 79 (3) ◽  
pp. 711-717 ◽  
Author(s):  
Dominique Lefier ◽  
Remy Grappin ◽  
Sylvie Pochet

Abstract The accuracy of fat, crude protein (CP), true protein (TP), and lactose determinations of raw milk by Fourier transform infrared (FTIR) spectroscopy and by analysis with a conventional filter-based milk analyzer was assessed in 6 trials performed over a 10-month period. At each trial, 30 bulk milk samples collected from 15 European countries and 11 reconstituted milks made from raw milk components were analyzed. When calibrations were performed with reconstituted milks at each trial, accuracy standard deviations for fat, CP, TP, and lactose were, respectively, 0.050,0.048,0.035, and 0.076 g/100 g for the filter instrument and 0.047, 0.046,0.042, and 0.065 g/100 g for the FTIR instrument. When a single calibration was made instead of calibrations at each trial, accuracy standard deviations increased for the filter instrument to 0.130, 0.119,0.121, and 0.083 for fat, CP, TP, and lactose, respectively, and for the FTIR instrument to 0.082, 0.053,0.044, and 0.084 g/100 g. Because the FTIR instrument provides more spectral information related to milk composition than does the filter instrument, single-calibration FTIR analysis of milk samples collected in different seasons is more accurate. Using reconstituted milks, prepared such that there is no correlation between fat, CP, and lactose, provides a more robust calibration than using genuine bulk milk, especially when milks with unusual composition are analyzed.


2020 ◽  
Vol 9 (3) ◽  
Author(s):  
Robin Dorau ◽  
Jun Chen ◽  
Peter Ruhdal Jensen ◽  
Christian Solem

The genome of Lactococcus lactis subsp. lactis bv. diacetylactis SD96, a strain used for cheese production, is presented. SD96 is refractory to phage attack, which is a desired property for starter bacteria. Its 10 plasmids provide industrially important traits, such as lactose and citrate metabolism, proteolytic activity, and phage resistance.


2008 ◽  
Vol 14 (6) ◽  
pp. 469-477 ◽  
Author(s):  
M. Dalmasso ◽  
S. Prestoz ◽  
V. Rigobello ◽  
Y. Demarigny

Cheeses are frequently made with natural whey starters (NWS). The whey from the previous cheese making is cultured and used for the next day. This practice, although essential for the development of typical sensory characteristics, can sometimes lead to acidification defects. In this work, the ability of Lactococcus lactis subsp. lactis biovar. diacetylactis to dominate over the other lactic acid bacteria (LAB) was tested in a controlled system as a possible explanation for these acidification breakdowns. A starter made of two Lc lactis subsp. lactis strains (LL), one Lc lactis subsp. cremoris strain (LC), and one Lc lactis subsp. lactis biovar. diacetylactis (LD) was added to sterilized milk. After incubation, the whey was removed and used to re-seed sterilized milk, the next day. This process was made during a five and twelve days' period. During the eight first days, the proportion of LD population increased, while the other LAB remained rather stable. Thereafter, LD strains dominated. At the same time, the diversity of LD population diminished considerably. If acidification ability of these LAB is not altered, this simplification was particularly hazardous in case of phages attack. LC and LL behavior was tested in milk containing increasing diacetyl concentrations. As long as diacetyl did not exceed a 5 ppm level — frequently measured in dairy products — its influence was impossible to detect. The selective advantage conferred by the citrate metabolism was proposed as a possible explanation for the LD population dominance. Other potential factors were also examined.


2021 ◽  
Vol 9 (6) ◽  
pp. 265-274
Author(s):  
Adna Bešić ◽  
Sead Karakaš ◽  
Kenan Čaklovica ◽  
Adisa Mušović ◽  
Mehmed Sultanović ◽  
...  

Motivation/Background: positive staphylococci (CPS) are common contaminants of raw milk. Before it is used, various heat treatments are applied to destroy microorganisms, inactivate enzymes and improve technological properties and concentration of dry matter of milk. This work aimed to determine the influence of commonly used heat treatments in diary on presence and number CPS in raw milk from Bosnia and Herzegovina area and to affirm whether there is a difference in efficacy between different treatments. Method: Using the standard method, 40 samples of raw milk from farms were inoculated for counting the initial number of CPS in raw milk. Samples were then exposed to heat treatments in vapor sterilizer and CPS number was counted using the same standard method. Results: Applied treatments included heat treatments at: 68 °C/40 s, 70 °C/15 s, 72 °C/without holding, 63 °C/30 min and 72 °C/15 s. CPS presence was detected in all tested samples of raw milk in numbers ranging from 2,82 to 5,32, with an average of 4,30, calculated as log10 /ml. Conclusions: Raw milk samples collected in the field initially registered a high CPS number. The applied heat treatments were effective to a large extent. The initial CPS count of milk seems to be the most important factor determining the number of CPS after heat treatments as well as traits of the strains.


2013 ◽  
Vol 56 (1) ◽  
pp. 709-718 ◽  
Author(s):  
L. Křížová ◽  
O. Hanuš ◽  
P. Roubal ◽  
J. Kučera ◽  
S. Hadrová

Abstract. The aim of the study was to describe the differences in nitrogen fractions and deepen the knowledge in the amino acid profile of raw milk affected by the breed of cattle, season and type of feeding. The study was conducted from June 2005 to February 2007 on 64 bulk milk samples collected from eight herds consisting of Czech Fleckvieh (four herds) and Holstein (four herds) breed. One half of the herds of each breed was grazed while the other half was not. Samples were collected twice in winter and twice in summer. The effect of the breed resulted in differences in milk yield that was lower in Czech Fleckvieh (5 385.50 kg) than in Holstein (7 015.15 kg, P<0.05). The content of nitrogen fractions was higher in Czech Fleckvieh than in Holstein (P<0.05). No effect of the breed on the amino acid profile of milk was observed except on the concentration of Glu (P<0.05). The effect of the season was demonstrated in the decrease of the concentrations of nitrogen fractions and Met during summer in comparison to winter (P<0.05). The effect of the type of feeding resulted in lower milk yield (5 197.50 and 7 203.75 kg) and lower concentrations of nitrogen fractions in grazed herds compared to non-grazed herds (P<0.05), respectively. Furthermore, the amino acid profile of milk differed significantly between grazed and non-grazed herds (P<0.05).


2014 ◽  
Vol 81 (4) ◽  
pp. 462-470 ◽  
Author(s):  
Georg Walcher ◽  
Monika Gonano ◽  
Judith Kümmel ◽  
Gary C. Barker ◽  
Karin Lebl ◽  
...  

Sampling approaches following the dairy chain, including microbiological hygiene status of critical processing steps and physicochemical parameters, contribute to our understanding of howStaphylococcus aureuscontamination risks can be minimised. Such a sampling approach was adopted in this study, together with rapid culture-independent quantification ofStaph. aureusto supplement standard microbiological methods. A regional cheese production chain, involving 18 farms, was sampled on two separate occasions. Overall, 51·4% of bulk milk samples were found to beStaph. aureuspositive, most of them (34·3%) at the limit of culture-based detection.Staph. aureuspositive samples >100 cfu/ml were recorded in 17·1% of bulk milk samples collected mainly during the sampling in November. A higher number ofStaph. aureuspositive bulk milk samples (94·3%) were detected after applying the culture-independent approach. A concentration effect ofStaph. aureuswas observed during curd processing.Staph. aureuswere not consistently detectable with cultural methods during the late ripening phase, but >100Staph. aureuscell equivalents (CE)/ml or g were quantifiable by the culture-independent approach until the end of ripening. Enterotoxin gene PCR and pulsed-field gel electrophoresis (PFGE) typing provided evidence that livestock adapted strains ofStaph. aureusmostly dominate the post processing level and substantiates the belief that animal hygiene plays a pivotal role in minimising the risk ofStaph. aureusassociated contamination in cheese making. Therefore, the actual data strongly support the need for additional sampling activities and recording of physicochemical parameters during semi-hard cheese-making and cheese ripening, to estimate the risk ofStaph. aureuscontamination before consumption.


1999 ◽  
Vol 62 (5) ◽  
pp. 547-548 ◽  
Author(s):  
LYNDA V. PODHORNIAK ◽  
SHANITA LEAKE ◽  
FRANK J. SCHENCK

Raw milk samples collected from bulk milk tankers may be screened for the presence of tetracycline antibiotics using rapid screening tests. If tetracycline residues are detected, the milk may be shipped to a laboratory for high-pressure liquid chromatography (HPLC) analysis. Because the milk may be shipped on ice blocks, it is important to know whether tetracycline residues are stable at that temperature and for how long. Control raw milk samples fortified with 50 ppb each chlortetracycline, demeclocycline, methacycline hydrochloride, minocycline, oxytetracycline, and tetracycline were incubated at 4°C or 25°C, then analyzed using a metal chelate affinity chromatography extraction and HPLC. No loss of tetracycline was observed after 48 h of storage at 4°C or 24 h at 25°C. Losses ranging from 4 to 13% and 0 to 18% were noted after 72 h at 4°C and 48 h at 25°C, respectively.


1977 ◽  
Vol 40 (1) ◽  
pp. 41-42
Author(s):  
EARL O. WRIGHT ◽  
WARREN S. CLARK ◽  
RICHARD W. WEBBER ◽  
WILLIAM L. ARLEDGE ◽  
MICHAEL H. ROMAN ◽  
...  

This collaborative study was done to determine whether laboratory personnel could successfully grade raw milk for sediment content using 0.40-, 0.20-, 0.14-, and 0.10-inch-diameter sediment discs. The 0.40-inch disc presently is accepted for grading sediment in mixed bulk milk samples. Technicians in 17 separate laboratories made 1,360 determinations (80 per laboratory) or 20 determinations for each size of sediment disc. Each laboratory graded the same set of samples. Samples were graded using photoprint standards that were prepared for this study. The laboratories were evaluated on the ability of laboratory technicians to grade the various sizes of sediment discs. Technicians in 13 of the 17 laboratories graded samples showing no significant differences in their ability to grade the various sized sediment discs. Four laboratories were not consistent in their ability to grade discs and showed significant differences in agreements with the previously determined sediment level as the diameter of the disc was reduced. Based on these results, most laboratories evaluated were capable of grading sediment discs with diameters of 0.40-, 0.20-, and 0.14-inch with consistency. With properly trained personnel in the laboratories, the study indicates the 0.10-inch diameter disc also can be used effectively for grading sediment in milk.


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