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Microbiological quality of bulk tank raw milk from two dairy farms in Hajdú-Bihar County, Hungary
105-112Views:327Two main channels have been identified to be responsible for microbiological contamination of raw milk and milk products. Firstly, contamination has occurred due to udder infection from the cow or the blood which harbours most bacteria that come in contact with the raw milk. Secondly, via external factors (may include faeces, skin, contaminated water, environment etc.) which are associated with the operation of milking. There is direct contact with the milk and/or surfaces before, during or after the milking, posing public health risk and economic decline. The aim of this study was to examine the bacteriological quality of bulk tank raw milk samples collected from two different size dairy farms (Farm 1 and Farm 2) of different housing forms (cubicle loose and deep litter) in Hajdú-Bihar County, Hungary in July, 2017. Three samples were taken from each farm, and the total plate count, coliform count, Escherichia coli count, Staphylococcus aureus count, and yeast and mould count were determined in them.
The results clearly showed low level of all measured bacteria group load in Farm 1 samples in comparison to Farm 2 with the exception of coagulase-negative Staphylococcus (CNS) which represented high level in general, indicating significant difference (P<0.05). The mean value of total plate count in Farm 2 samples was higher (1.0 × 105 CFU/mL) than Farm 1 samples (2.8 × 104 CFU/mL). There was a significant difference (P<0.05) in mean count of coliforms in raw milk samples between Farm 1 and Farm 2. Similarly, results of E. coli were significantly different (P<0.05) with mean count of 1.44 × 102 CFU/mL and 2.02 × 103 CFU/mL for Farm 1 and Farm 2 respectively.
Results of Staphylococcus aureus also showed significant difference (P<0.05) with mean count of 9.7 × 101 CFU/mL for Farm 1 and 6.28 × 102 CFU/mL for Farm 2. The mean of mould count recorded was 1.07 × 102 CFU/mL and 4.93 × 102 CFU/mL for Farm 1 and Farm 2 respectively. The recorded mean of yeast count was 1.68 × 103 CFU/mL and 3.41 × 103 CFU/mL for Farm 1 and Farm 2 respectively; however, both farms showed no significant difference (P>0.05) in terms of mean of mould and yeast count. Although Farm 2 produced six times lower milk quantity than Farm 1, the measured microbial parameters were high. Both farms’ microbiological numbers were higher above the permitted limit values as stated by Regulation (EC) No 853/2004, Hungarian Ministry of Health (MoH) 4/1998 (XI. 11.).
This could be an indication of non-conformance to effective GMP, ineffective pre–milking disinfection or udder preparation, poor handling and storage practice, time and temperature abuse and inadequate Food Safety Management System Implementation. Therefore, our recommendation is as follows; establish control measures for pre- and postharvest activities involved in the milking process which would be an effective approach to reduce contamination of the raw milk by pathogenic microorganisms from these farms, strict sanitation regime and hygiene protocol be employed and applied to cows, all equipment, contact surfaces and minimize handling of the milk prior, during and after milking. This will also serve as scientific information to the producers for continual improvement in their operations.
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Microbiological status of bulk tank milk and different flavored gomolya cheeses produced by a milk producing and processing plant
73-78Views:275The microbiological quality of milk is important not only for food safety, but it can also influence the quality of dairy products. In this study, our aim was to assess the microbiological status of the bulk milk of a milk-producing farm, and some natural and flavored (garlic, dill, onion) gomolya cheeses made from pasteurized milk produced by their own processing plant. We determined the number of coliform bacteria, Escherichia coli, Staphylococcus aureus, and molds of three milk and eight cheese samples. The tests were conducted between July and September, 2017.
In bulk milk, the mean coliform count was 3.83±0.17 log10 CFU/ml; the mean E. coli count was 1.38±0.14 log10 CFU/ml; the mean mold count was 3.74±1.30 log10 CFU/ml; and the S. aureus count was <1.00 log10 CFU/ml, respectively. The mean coliform count in gomolya cheeses was 3.69±1.00 log10 CFU/g; the mean E. coli count was 2.63±0.58 log10 CFU/g; the mean S. aureus count was 3.69±1.35 log10 CFU/g and the mean mold count was 1.74±0.37 log10 CFU/g. The amount of coliforms detected in different flavored gomolya cheeses were significantly different (P<0.05). More than 10 CFU/g of E. coli was found only in the dill flavored cheeses, and S. aureus was found only in dill (3.66±1.86 log10 CFU/g) and onion (3.71±0.52 log10 CFU/g) flavored gomolya cheeses. Based on the obtained results, it was found that the amount of coliform bacteria and E. coli in bulk milk exceeded the limit set in regulation of the Hungarian Ministry of Health (MoH) 4/1998 (XI. 11.) and the amount of S. aureus was below the limit. For gomolya cheeses, the S. aureus count exceeded the limit. The amount of coliform bacteria remained above the limit in cheeses, except for the garlic flavored gomolya cheese. In cheeses, a larger E. coli count was detected than in the bulk milk, but there is no specific limit for cheeses in the regulation. The mold count exceeded the limit specified in the regulation in cheeses, but a lower value was detected relative to milk.
The results show that, in the case of bulk milk and gomolya cheeses, certain detected quantities exceeded the limit values set forth in regulation of MoH 4/1998 (XI. 11.). The results indicate an inadequate microbiological state of the raw material and the finished products. The reasons for these are due to reduced technological hygiene or the inappropriate handling of raw material and finished products. In this study, we have summarized the results of our preliminary studies, which can provide a basis for further hygiene studies.
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Isolation and identification of endophytic fungi connected to Grapevine Diseases, from the Tokaj wine region, Hungary
61-66Views:224Grapevine Trunk Diseases (GTD) is one of the most important diseases in vineyards worldwide, which can be found in Hungarian vineyards as well. In Hungarian wine regions there is very little information about the occurrence of pathogens which cause GTD, in case of Tokaj wine region there is no knowledge about that, what kind of pathogens can be found in the vineyards.
The objective of our research is to assess the situation and occurrence of GTD in Tokaj wine region in cooperation with local specialists, as well as identification of pathogens which were isolated from the diseased trunks by morphological and genetic basis.
We were able to isolate endophytic fungi from all sampled grape trunk. The majority of them were determined as Diplodia seriata not only based on colony morphology, but also determined by rDNA sequences.
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Definition antioxidant activity of selenium-enriched food sprout, as well as their microbiological analysis
25-30Views:255In this present study, we prepared selenium-enriched food sprouts, where the antioxidant capacity was analysed, we also determined their microbiological status. We took into account the fact, we choose micronutrients to our treatment, that selenium can be delivered to the body by a small amount with the most widely consumed food.
We focused during our research to determine that the increasing concentrations of selenium treatment, in which we used sprouts, knowing fully well that it has an impact on aboriginal antioxidant capacity of sprouts, which is mainly due to high vitamin content of sprouts.
Furthermore, we think it is important to make microbiological analysis, because germination conditions, for example temperature, pH, all this will create an ideal environment for the growth of microorganisms. So we had goal to determine, how the used selenium concentration affect the total plate count, coliform bacteria count and Staphylococcus aureus count of sprouts.
We determined the aboriginal water-soluble and lipid-soluble antioxidant capacity of sprout with the PHOTOCHEM chemiluminometer and we applied pour plate technique for the mapping of the mycrobiological state of sprouts.
Experimental results are evaluated, we state that increasing concentrations of selenite or selenate treatment, is primarily water-soluble antioxidant capacity of sprouts was affected. The water-soluble antioxidant capacity of wheat sprout was much higher than the measured values in pea sprout, this may be linked to what we measured. That is much higher ascorbic acid content in case of wheat sprout, which is well known as one of the most important antioxidant properties compounds of wheat sprout.
We conclude from the results of the microbiological, that the highest concentrations of selenite or selenate treatment has a relative significant anti-microbial effect in case of wheat sprouts. Coliform and total plate count showed no clear decreasing tendency, although the values of treatments in both cases obtained were below the control values.
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Comparative analysis of Staphylococcus aureus strains by molecular microbiology methods
34-39Views:167Staphylococcus aureus is a very important pathogen for dairy farms and milk processing plants. Subclinical mastitis is often caused by this species, and it can contaminate bulk tank milk when milking cows are suffering from mastitis. Additionally, thermostable enterotoxins (SE) produced by some types of this bacterium can cause food poisoning.
The aim of our research was to examine the number of S. aureus in bulk tank milk in two dairy farms and the enterotoxin-producing ability, genetic relation (pulsotype) and antibiotic resistance of S. aureus strains from different sources (bulk tank milk, udder quarter milk and environment).
The results show that the mean number of S. aureus of bulk tank milk of two farms significantly differed (P<0.05). Fourteen isolates were selected for further molecular genetic studies (five isolates were from bulk tank milk and nine isolates were from udder quarter milk). S. aureus was not recovered from the environmental samples. Three of the fourteen isolates (21.4%) tested by multiplex PCR were positive for SE genes. Two isolates carried one gene (seb) and one isolate carried two genes (seg and sei). The fourteen strains were classified into three pulsotypes and two subtypes at 86% similarity level. Isolates from bulk tank milk (n=5), were divided into 2 pulsotypes (A, C) and one subtype (C1). The isolates from udder quarter milk (n=9) belonged to three different pulsotypes (A, B, C) and two subtypes (A1, C1). The distribution of pulsotypes in the present study revealed genetic relationship between S. aureus isolated from udder quarter milk and bulk tank milk. This could be explained by the fact that in farms with a high number of infected cows, these cows could represent the main source of contamination. The results of the antibiotic resistance investigations show, that all strains were susceptible to methicillin, cefoxitin, lincomycin, tetracycline, erythromycin and sulfamethoxazole/trimethoprim. Thirteen out of fourteen strains were resistant to penicillin (A and C pulsotypes, A1 and C1 subtypes) and just one isolate was susceptible (B pulsotype) to all antibiotics tested. -
Bacteria in the milk of sheep with or without mastitis- mini Review
47-52Views:373From a nutritional point of view, sheep milk is more valuable than cow and goat milk and the interest for sheep milk is increasing in many countries. However, sheep milk is easily contaminated during milking, handling, and transport and it is an ideal medium for bacterial propagation. Consequently, sheep milk spoils quite quickly. The proper, clean handling of milk is not only of sanitarian interest, but it also serves the farmers’ interests, because contaminated milk may not be distributed, and is unsuitable for producing good quality products. Following this technological trend, this review addresses the bacterial composition of sheep milk with and without mastitis. Even though sheep milk contains a lot of bacteria, this review article highlighted total plate count, Enterobacteriaceae, coliform, Staphylococcus aureus, Escherichia coli, Listeria monocytogenes, Campylobacter, Salmonella spp. and Streptococcus spp. Mastitis in sheep is a vital cause of mortality, reduction in milk production and early culling. The reported risk factors for mastitis in sheep were age, a case of mastitis, breed, husbandry systems, and location. The main priority should be implementation of programs to minimize human pathogenic bacteria and mastitis in raw ewe milk.
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The effect of keeping technology on the microbiological status of raw milk
67-75Views:135The importance of the quality of raw milk increased after Hungary had joined to the EU. On delivery of raw milk, the microbiological quality, especially total plate count of the milk is very important. Twenty-two farms (7 large, 4 medium-sized, and 11 small farms) were included in the study. We considered the different farm size, keeping- and milking circumstances during the selection of farms. The examined large farms use loose housing system (cubicle, deep litter) and milking parlour. Most of them use preand post-milking disinfection. In the medium-sized farms, loose,
deep litter and tie-stall housing system, as well as milking parlour, pipeline milking and bucket milking occurred. All of them use preand post-milking disinfection. Small farms use tie-stall housing system, bucket milking and udder preparation by water. Unfortunately, they do not use pre- or post-milking disinfection. In the large and medium-sized farms mainly Holstein Friesian, in the small farms Hungarian Simmental breeds can be found.
The aim of our research was to examine the microbiological status of the raw milk produced in dairy farms (total plate count, coliform count, Escherichia coli count, Staphylococcus aureus count, psychrotroph bacteria count, furthermore yeast and mold count); sources of the contamination; connection between the microbiological quality of produced milk and housing-, milking technologies of farms; furthermore the hygienic circumstances of milking and milk handling of the farms, by the examination of coliform bacteria and Escherichia coli contamination.
During the examination of the connection between the different farm sizes, various housing- and milking forms and the microbiological characteristics we observed similar tendencies in the case of total plate count, coliform count, yeast and molds count, furthermore psychrotroph bacteria count. The value of these parameters was significantly higher in small farms, and infarms which use tie-stall housing forms, bucket milking, udder preparation with water, and which do not use pre- and post-milking disinfection.
The results showed that besides cooling, the milking procedure and the type of udder preparation had the largest effect on the total plate count. Statistical analysis shows that in medium and small farms the combination of pipeline milking – tie stall housing system – disinfectant preparation of the udder; in large farms the combination of milking parlour – loose cubicle housing system – dry preparation of the udder are the most appropriate in the aspect of the total plate count. We experienced that in farms where the hygienic instructions are not followed – and therefore
equipment used during the milking and handling of milk is very contaminated – or rather the separation of mastitic cows’ milk is not appropriate, different microorganisms may contaminate the produced milk. -
Coagulase-negative staphylococci in ewe udder surface and raw milk samples
221-225Views:181Coagulase-negative staphylococci (CNS) are among the major causes of subclinical mastitis in dairy ewe flocks. This has a financially significant impact on the ewe dairy sector and consumer health. The determination of the bacterial count, particularly CNS, is critical in terms of the quantity and quality of ovine milk. Thus, the purpose of this study was to quantify the CNS count in the udder surface and raw milk samples of the ewe, in addition to identifying CNS strains (n=8) collected from udder surface and individual raw milk samples by 16S rRNA gene sequencing. A total of 164 samples of udder surface and raw milk originated from four sheep farms were tested. The obtained values were compared between the different farms. Furthermore, values during 2018 and 2019 in the case of Farm I were compared. Significantly higher (p<0.05) average count was observed in udder surface samples taken from Farm I (2.8±1.0 lg CFU/cm2) than that of Farm III samples (2.3±0.6 lg CFU/cm2). In the case of individual raw milk, the higher value was observed in samples derived from Farm III (3.5±0.9 lg CFU/mL), while the obtained value was significantly lower (p<0.05) in samples originated from Farm IV (1.8±0.4 lg CFU/mL). In the bulk tank milk samples, the highest mean value was 5.3±0.4 CFU/mL, and there was no significant (p>0.05) variation between farms. Coagulase-negative staphylococci counts were decreased to a certain extent in both sample types tested during 2019 except for individual raw milk derived from the Tsigai breed. The correlation between the mean CNS counts of udder surface and individual raw milk was very weak (r=0.048). Staphylococcus simulans, Staphylococcus auricularis, and Staphylococcus equorum were identified by molecular sequencing and Staphylococcus simulans were the most frequently identified CNS species. A higher CNS count of bulk tank milk than individual raw milk indicates possible contamination during milking and storage. Therefore, further studies are required to investigate the other sources of bulk tank milk contamination to improve the hygienic quality of milk.
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The effect of breed and stage of lactation on the microbiological status of raw milk
37-45Views:339The microbiological quality of the milk is important not only for food safety, but it can also influence the quality of dairy products. The microbiological status of raw cow milk can be influenced by many factors. Our aim was to determine whether there was a difference between the microbiological quality of milk of two different cow breeds (Holstein Friesian and Jersey) kept and milked in the same conditions, and how the microbiological quality of the raw cow milk changed during lactation (beginning, mid, and end). Samples were taken and analysed in July, August and September in 2018 from two dairy farms in Hajdú-Bihar county. During the conducted studies, the total plate count (TPC), the coliform count, the Staphylococcus aureus count and the coagulase-negative Staphylococcus (CNS) count of raw milk samples were determined.
There was no significant difference (P>0.05) between the milk of the Holstein Friesian and Jersey breeds in the case of TPC. However, the mean coliform count of milk samples taken from Holstein Friesian cows was significantly lower (P<0.05) than the mean coliform count of milk samples taken from Jersey cows. S. aureus was detected in one of the twelve milk samples taken from Holstein Friesian cows, and in two of the eleven milk samples taken from Jersey cows. CNS was found in larger amount in milk samples taken from Holstein Friesian cows, and the difference was significant (P<0.05). Both TPC and CNS count were significantly higher (P<0.05) in individual milk samples taken at the end stage of lactation, than in samples taken in the earlier stages of lactation from Farm “A”. However, in the case of Farm “B”, there was no significant difference (P>0.05) in colony counts at different stages of lactation. S. aureus was only present in milk samples that collected from cows, which were at the beginning and middle stages of lactation. Testimg the hemolysin production ability of S. aureus strains isolated from the raw milk samples, only weak hemolysis was observed on blood agar. In case of antibiotic resistance testing, it was found that all strains were susceptible to cefoxitin, chloramphenicol, clindamycin, erythromycin, gentamicin, penicillin G, tetracycline and trimethoprim/sulphamethoxazole.
Based on the results of our studies, staphylococci were detected in a higher amount in the milk of Holstein Friesian cows, and coliform bacteria were detected in a higher number in the milk of Jersey cows. Summing up the results of the milk samples taken from the different stages of lactation in one of the farms, it can be concluded that higher TPC and CNS count could be detected at the end stage of lactation than in the samples taken from the earlier stages of lactation. The fact that at the end of lactation the microorganisms could be detected in a higher colony count may be related to the fact that teats could be damaged during lactation by the milking machine, which increased the chance of imvading the microorganisms into the udder.
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Comparison of sample preparation methods for the identification of Staphylococcus Aureus by MALDI-FOF MS
9-14Views:298Coagulase-positive staphylococci include 3 species, Staphylococcus aureus, S. hyicus and S. intermedius. Of these three species, S. aureus is the most well-known human pathogen. S. aureus is part of the human and animal normal microbiota, however, it is capable of producing several staphylococcal enterotoxins (SEs) that cause intoxication symptoms of varying intensity in humans after consuming contaminated food. Selective media which are used for the determination of coagulase-positive staphylococci from foods are not able to identify isolates at a species. With the MALDI-TOF MS technique, we can identify S. aureus cheaper and faster than by using molecular methods. This paper describes the results of the study of the presence of coagulase-positive staphylococci and S. aureus in many food products, and the application of three sample preparation methods: direct sample preparation, formic acid suspension and ethanol extraction.
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The examination of presumed Escherichia coli count of raw milk samples on several milk production farms
31-37Views:172For dairy farms, it is of great importance to insure the appropriate hygienic status of milk and to examine it regularly. Escherichia coli, belonging to the coliform bacteria type of, is a good indicator of contamination, and therefore suitable for characterising the hygienic condition of milk production.
The aim of our research was to examine the connection between the Escherichi coli count in bulk tank milk and housing and milking technologies of different-sizes farms. We examined the relation using various statistical methods.
Analysing the connection between the E. coli count and the farm size we found no significant difference between the farms. On the basis of the mean values of the E. coli count, we can say that the hygienic conditions are appropriate for mid-sized farms, and tolerable for large farms. We found differences in the hygienic status among the small farms. Half of the eight small farms, had no adequate hygiene. The results of the analysis of the quality categories show that the probability of inadequate quality milk was the largest on small farms (37.5%).
Comparing the various housing and milking methods with each other, there were numerical differences in the E. coli count, but these differences were not significant. We got higher E. coli count values on those farms using tied stall barn and bucket milking installation. The reason for this could be that in cases of farms using bucket milking installation, it is harder to meet the requirements.
After forming groups by farm size, housing and milking methods, we found that the E. coli counts are adequate on mid-size farms using various housing and milking methods; and tolerable on those large farms using loose housing stable and a milking parlour. At the same time, we found inadequate E. coli counts on the smaller farms using tied stall barns and bucket milking installation.
The results show that if there is suitable attention, independent of farm size, housing and milking procedure, it is possible to produce milk with low E. coli counts, and to insure appropriate hygienic conditions.
Further detailed examinations are needed to decide which factors of housing and milking technologies influence the E. coli count of bulk tank milk. -
The effect of sous-vide cooking on the antioxidant properties of oyster mushroom (Pleurotus ostreatus L.)
177-184Views:165Oyster mushrooms (Pleurotus ostreatus L.) are renowned for their antioxidant, antimicrobial, and prebiotic properties. This study explores the antioxidant characteristics, activity, and β-glucan content in freeze-dried mushroom samples, investigating the influence of sous-vide cooking. Uncooked freeze-dried P. ostreatus and three pre-cooked freeze-dried samples (70, 80, 90 °C through 4 hours) were analysed for Total Polyphenol Content (TPC), Total Flavonoid Content (TFC), Radical Scavenging (DPPH), Ferric Reducing Antioxidant Power (FRAP), and β-glucans content via HPLC and Total Dietary Fiber (TDF) via enzymatic gravimetric method. Results indicate that uncooked mushroom powder exhibited superior antioxidant capabilities compared to cooked samples. The sous-vide cooked (80 °C) mushrooms displayed the highest total phenolic and flavonoid content. Moreover, pre-cooked (70 °C) mushroom powder demonstrated the highest β-glucan content, significantly surpassing the uncooked control sample. Notably, pre-cooked groups (80, 90 °C) demonstrated significantly higher TDF levels compared to uncooked sample. This research offers valuable insights into the potential use of mushrooms as high-antioxidant, antimicrobial, and prebiotic food or feed supplements, with broad implications across various fields.
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The effect of season on the microbiological status of raw milk
95-99Views:181Many factors can influence the microbiological quality of raw cow’s milk. In this study, our aim was to determine whether there was any difference between the microbiological statuses of milk produced in different seasons. Samples were collected and analysed from five dairy farms in Hajdú-Bihar County, from February to November in 2019. During our studies, total plate count (TPC), coliform count and Staphylococcus aureus count of raw cow’s milk samples were determined.
There was no significant difference (P>0.05) between the mean TPC values detected in the milk collected in winter and autumn, but that values were significantly (P<0.05) lower than in the milk samples collected in spring and summer. Similarly to the TPC, in the case of coliform bacteria the lowest mean colony count was detected in the samples collected in winter. The difference was significant (P<0.05), compared to the values observed in the samples collected in summer. S. aureus was detected in bulk milk of only two farms in excess of 1.0 log10 cfu/ml. Also in case of S. aureus, there was a significant difference (P<0.05) between the values observed in the samples collected in winter and in summer. Samples from spring and summer contained the highest amount of S. aureus.
Based on the results of our studies, in the case of almost all farms the mean TPC, coliform and S. aureus counts were lower in the samples collected in winter, than in the samples collected in summer. The fact that the samples collected in winter contained the lowest amount of colonies could be attributed to the inhibition of growth of mesophilic microorganisms below 8 °C. Furthermore, the fact that we observed the highest colony counts in samples collected in summer, can be related to the heat stress of cows during the summer due to unfavorable weather conditions (high temperature and humidity).
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The factors affecting the proliferation of mould fungi and mycotoxin production during the storage of wheat and the identification methods of the appearing branches
129-133Views:142Nowadays, it is often suggested that, we should eat products made with whole grain cereals, despite of the fact that it raises the risk of consuming wheat products infected by mold and their toxins originated from the plough-lands and the stocks.
Two third of the cultivated fields in Hungary are planted with cereals. The most alarming problem for food and feed security is caused by the Fusarium species. The greatest problem of all is caused by the mycotoxins. When they get into the food chain they can be a serious threat to public health. In addition, we have to face up to the problem of the effects of global warming that influence the growth of microbial infections in different ways.
In this article we tried to summarize the effect of climate change on molds, the factors which have effect on growing and mycotoxin producing of molds and the identification methods of molds.