MYCOLOGICAL EVALUATION OF MILK AND SOME MILK PRODUCTS IN BENI-SUEF CITY

Document Type : Research article

Authors

Faculty of Veterinary Medicine, Beni Suef University, Beni-Suef, Egypt

Abstract

A total of 125 random samples from raw milk, locally manufactured kareish cheese, hard cheese (Rass cheese), plain yoghurt and flavored yoghurt (25 of each) collected from dairy shops and super markets in Beni Suef city were subjected to mycological examination. The yeasts were detected in milk, Rass cheese and locally manufactured kareish samples with a mean value of 6.22Χ102 ± 3.62Χ102, 14±7.21 and 3.23Χ106 ± 2.017Χ106 respectively, while plain yoghurt and flavored yoghurt were free from any yeasts (<10). The most predominant isolated spp. were Candida pseudotropicalis, Rhodotorula species, Candida species, Torulopsis (candida) glabrata and Cryptococcus neoformans. The mean value of molds was 2.23Χ102±9.3Χ101, 4.8±1.54, 1.068Χ106±7.20Χ105, and 2.4±0.871 in the examined milk, Rass, locally manufactured kareish cheese and flavored yoghurt samples respectively, while molds couldnt be detected in the examined plain yoghurt samples. The most predominant isolated spp. Were Mucorspp, Asperigellus flavus, Asperigellus niger, Asperigellus fumigatus, Absidia spp., Penicillium spp., Geotrachum candidum, Cladosporium spp., Phoma spp. and Fusarium spp < /em>. All the examined Rass cheese samples were free from aflatoxin M1 calibrated by fluorometer. The economic and public health significance of the fungi and mycotoxins as well as the sanitary precautions were mentioned.

Keywords


Assiut University web-site: www.aun.edu.eg

 

MYCOLOGICAL EVALUATION OF MILK AND SOME MILK PRODUCTS IN

BENI-SUEF CITY

 

SAADIA H. EL-SHINAWY; ADEL M. EL-KHOLY; ARAFA M.S. MESHREF and

SAMMER W. SHARKAWY

 

Faculty of Veterinary Medicine, Beni Suef University, Beni-Suef, Egypt

 

Received: 31 December 2017;       Accepted: 30 January 2018

 

ABSTRACT

 

A total of 125 random samples from raw milk, locally manufactured kareish cheese, hard cheese (Rass cheese), plain yoghurt and flavored yoghurt (25 of each) collected from dairy shops and super markets in Beni Suef city were subjected to mycological examination. The yeasts were detected in milk, Rass cheese and locally manufactured kareish samples with a mean value of 6.22Χ102 ± 3.62Χ102, 14±7.21 and 3.23Χ106 ± 2.017Χ106 respectively, while plain yoghurt and flavored yoghurt were free from any yeasts (<10). The most predominant isolated spp. were Candida pseudotropicalis, Rhodotorula species, Candida species, Torulopsis (candida) glabrata and Cryptococcus neoformans. The mean value of molds was 2.23Χ102±9.3Χ101, 4.8±1.54, 1.068Χ106±7.20Χ105, and 2.4±0.871 in the examined milk, Rass, locally manufactured kareish cheese and flavored yoghurt samples respectively, while molds couldnt be detected in the examined plain yoghurt samples. The most predominant isolated spp. Were Mucorspp, Asperigellus flavus, Asperigellus niger, Asperigellus fumigatus, Absidia spp., Penicillium spp., Geotrachum candidum, Cladosporium spp., Phoma spp. and Fusarium spp. All the examined Rass cheese samples were free from aflatoxin M1 calibrated by fluorometer. The economic and public health significance of the fungi and mycotoxins as well as the sanitary precautions were mentioned.

 

Key words: Yeast, mold, mycotoxins, milk, dairy products.

 

 


INTRODUCTION

 

Milk has been used by human since the beginning of the life  as the mostpopular and nutritious safe human food, this value put the food hygienists in a real challenge, to provide safe  milk to consumer and at the same time keep its nutritional value high as much as possible. In some countries including Egypt, most milk is manufactured into more stable dairy products of worldwide commerce, such as cheese andyoghurt.

 

The manufactured dairy products from milk have the same properties of original milk moreover; several stages of processing may be unfavorable and add more points of hazard and weakness allowing entrance of molds and unfortunately, these products support molds growth and toxins production due to its contents of protein, low pH and storage under unfavorable conditions Gqaleni et al. (1997); Pardo et al. (2004).

 

 


Corresponding author: Dr. SAMMER W. SHARKAWY

E-mail address:ashrykhaled@gmail.com

Present address: Faculty of Veterinary Medicine, Beni Suef University, Beni-Suef, Egypt

Yeasts and molds can grow in milk and its products particulary at suitable conditions of temperature and moisture Barrois et al. (1997). Contamination of these products may occur from the raw material or during the manufacturing, storage and distribution Kure et al. (2004). Such microorganisms influence the biochemical characters and flavor of such products as well as their appearance rendering them commercially undesirable and often resulting in decreasing the grading of the dairy products Muir and Banks (2000).

 

Mycotoxins produced by certain molds as a toxic substances can be found in dairy products from two origins: indirect contamination, which results when dairy cows ingest feed that contains mycotoxins which pass into the milk such as aflatoxin M1 and direct contamination, which occurs due to the intentional or accidental growth of molds on the surface of the dairy products and  secrete aflatoxins B1, B2, G1 and G2 Sengun et al. (2008) therefore, feeding on a low quality ration contaminated with molds more than 10 cfu /g and kept under humid conditions cause intoxication to both animals and human whom consumed this animal products.

 

The present work was performed to investigate the presence of molds and yeasts in milk and some milk products in Beni- Suef city, identification of isolated strains and detection of mycotoxins in Rass cheese samples.

 

MATERIALS AND METHODS

 

Collection of the samples:

A total of 125 random samples from raw milk, locally manufactured kareish cheese, hard cheese (Rass cheese), plain yoghurt and flavored yoghurt (25 of each) were collected from dairy shops and super markets in Beni-Suef city for mycological and mycotoxicological examination.

 

Preparation of the samples (A.P.H.A.1992):

Raw milk samples were subjected to storch's test lampert (1975) to exclude all samples proved to be heat treated.

 

Rass and kareish cheese samples were thoroughly mashed before being emulsified in the diluted solution.

 

Whole yoghurt samples were perfectly mixed before examination.

Eleven ml or g of each prepared fluid milk and yoghurt samples were added to 99ml of sterile saline to make adilution of 1/10, from which serial dilutions were prepared. In Rass cheese and kareish cheese samples, Eleven grams of prepared samples were aseptically transferred to a sterile blender jar and 99 ml of warmed (40°C) sterile 2% sodium citrate solution were added and the contents were blended till completely emulsified to make a dilution of 1/10 from which decimal dilution were prepared.

 

Mycological examination (Bailey and Scott, 1978):

From the already prepared serial dilution, one ml was transferred into duplicate petri-dishes and thoroughly mixed with 15 ml of sabaroud dextrose agar containing 50 ppm chloramphenicol Hup and Staddhouders (1972). The inoculated plates were incubated at 25°C for 7 days. The first examination of the plates was done after 3 days to determine the degree of fungi growth. The yeasts and molds count /ml or g was calculated and recorded.

 

Representative colonies were isolated on the sabaroud dextrose agar slopes which incubated at 25°C for 5 days for further identification.

 

Isolated molds were identified according to Ramirez (1982); Samson (1979), while yeasts colonies were identified according to lodder (1970).

 

Aflatoxin M1 in Rass cheese samples was detected by using fluorometerScott and Trucksess, (1997).

 

 

RESULTS

 

Table 1: Statistical analytical results of yeasts and molds Counts in the examined raw milk samples.

Isolates

No of the examined

samples

No of positive samples

Min/ml

Max/ml

Mean/ml

± SEM

No

%

Yeasts

 

25

17

68

<10

9Χ103

 

6.22 Χ102

3.62 Χ102

Molds

 

25

16

64

<10

1.6Χ103

2.23Χ102

9.3Χ101

 

Table 2: Statistical analytical results of yeasts and molds counts in the examined Rass cheese samples.

 

Isolates

No of the examined

samples

No of positive samples

Min/g

Max/g

Mean/g

± SEM

No

%

Yeasts

 

25

6

24

<10

1.5Χ102

14

7.21

Molds

 

25

8

32

<10

0.2Χ102

4.8

1.54

 

Table 3: Statistical analytical results of yeasts and molds counts in the examined Kareish cheese samples.

 

Isolates

No of the examined

samples

No of positive samples

Min/g

Max/g

Mean/g

± SEM

No

%

Yeasts

 

25

16

64%

<10

0.5Χ108

 

3.23Χ106

2.017Χ106

Molds

 

25

15

60%

<10

1.8Χ107

1.068Χ106

7.20 Χ105

 

Table 4: Statistical analytical results of yeasts and molds counts in the examined large scale yoghurt samples.

 

The examined samples

Isolates

No of the examined

samples

No of positive samples

Min/g

Max/g

Mean/g

± SEM

No

%

Flavoured yoghurt

Yeasts

 

25

0

0

<10

<10

<10

<10

Molds

 

25

6

24

<10

0.1Χ102

2.4

0.87

Plain yoghurt

Yeasts

 

25

0

0

<10

<10

<10

<10

Molds

 

25

0

0

<10

<10

<10

<10

 

Table 5: Incidenceof the identified yeasts isolates.

 

Yeast species

 

 

 

No of the examined

samples

Milk

Kareish cheese

Romano cheese

Plan yoghurt

Flavoured yoghurt

+ve

%

+ve

%

+ve

%

+ve

%

+ve

%

Candida pseudotropicalis

25

6

24%

-

-

1

4%

-

-

-

-

Rhodotorula species

25

1

4%

-

-

-

-

-

-

-

-

Candida species

25

9

36%

12

48%

3

12%

-

-

-

-

Torulopsis (candida) glabrata

25

1

4%

3

12%

2

8%

-

-

-

-

Cryptococcus neoformans

25

-

-

2

8%

-

-

-

-

-

-

 

 

Table 6: Incidenceof the identified molds isolates.

 

Molds species

 

 

 

No of the examined

samples

Milk

Kareish cheese

Romano cheese

Plan yoghurt

Flavoured yoghurt

+ve

%

+ve

%

+ve

%

+ve

%

+ve

%

Mucor spp.

25

2

8%

-

-

2

8%

-

-

2

8%

Asperigellus

flavus

25

2

8%

6

24%

1

4%

-

-

2

8%

Asperigellus niger

25

3

12%

7

28%

2

8%

-

-

-

 

Asperigellus fumigatus

25

1

4%

-

-

2

8%

-

-

-

 

Absidia spp.

25

-

-

2

8%

1

4%

-

-

-

 

Penicillium spp.

25

-

-

-

-

-

 

-

-

1

4%

Geotrachum candidum

25

6

24%

-

-

-

 

-

-

1

4%

Cladosporium spp.

25

1

4%

-

-

-

 

-

-

-

 

Phoma spp.

25

1

4%

-

-

-

 

-

-

-

 

Fusarium spp.

25

1

4%

-

-

-

 

-

-

-

 

 


DISCUSSION

 

The Results given in Table 1 revealed that the examined milk samples were contaminated with yeasts and molds at percentages of 68% and 64%, respectively with a mean value of 6.22Χ102± 3.62Χ102cfu/g and 2.23Χ102±9.3Χ101 cfu/g, respectively. Similar results were reported by Ghazal  (2001), while higher molds and yeasts counts were observed in samples examined by El-Diasty and El- Kaseh (2009); Pešić-Mikulec et al. (2005); Lavoie et al. (2012). The obtained results indicated poor hygiene during handling which might lead to technological problems during processing.

 

The data summarized in Table 2 revealed that yeasts and molds were isolated from Rass cheese samples at a percentage of 24% and 32% respectively with a mean value of 14±7.21cfu/g and 4.8±1.54cfu/g respectively. Higher counts with lower incidence were reported by Al- Hawary et al. (2002); Amer (2002); El-Asuoty (2011).

 

The obtained results in Table 3 showed that the count of total yeasts and molds in the examined kareish cheese samples were detected at a percentage of 64% and 60% respectively with a mean value of 3.23×106± 2.017×106 and 1.068×106± 7.20×105 cfu/g. Similar counts were detected by Moawad et al. (2002), while higher counts with lower incidence were reported by Khair-Allah (2000); Hassan (2003), lower counts and incidence were recorded by El-shrief (2000); El-Komy (2002).

 

The results given in Table 4 proved that the total molds existed in the examined flavored yoghurt samples at a percentage of 24% with a mean value of 2.4 ± 0.87178cfu/g. Higher results for the count were recorded by Moustafa (2004);  Hafez (2010); Hassan (2003).

 

On the other hand the yeasts failed to be detected in the flavoured yoghurt samples, but both yeasts and molds couldn't be detected in the plain yoghurt samples. The achieved results allow concluding that most of the examined samples proved to be not contaminated with molds and yeasts, which indicate good hygienic measures during production, handling, storage and distribution. The presence of molds in large scale products may be attributed to post pasteurization contamination and/or unsanitary measures during handling and distribution.

 

Species of Candida pseudotropicalis, Rhodotorula species, Candida species, Torulopsis (candida) glabrata and Cryptococcus neoformans could be isolated from the different samples at varying percentage in Table (5). Several authors obtained the same results as El-Diasty and El- Kaseh (2009); Sarais et al. (1996).

 

Species of Mucor spp., Asperigellus flavus, Asperigellus niger, Asperigellus fumigatus, Absidia spp., Penicillium spp., Geotrachum candidum, Cladosporium spp., Phoma spp. and fusarium spp. could be isolated from the perivous samples as in Table 6 Several authors obtained the same results El-Diasty and El- Kaseh (2009); Karthikeyan and Pandiyan (2013) and Gulbeand Valdovska (2014).

 

On the other hand aflatoxin M1 residue couldnt be detected in the examined Rass cheese samples. The result is similar to Robinson and tamime (1991); Martins et al. (1995); Škrinjar et al. (2011).

 

Mold and yeast contamination not only causes deterioration of food but also can adversely affect the health of humans. Moreover, fungi influence the biochemical characters and flavor of the product and its appearance is commercially undesirable and often result in down grading of the product.

 

It is concluded that strict hygienic measures should be applied during production, processing and distribution of milk and its products to avoid contamination. Periodical inspection must be done by specialists on the dairy farms to minimize milk contamination with different types of yeasts and molds. The milk obtained from dairy animals fed on feedstuffs contaminated with aflatoxin B1 must be rejected. Efficient cleaning and sanitization of farm dairy utensils must be done to improve the quality of raw milk and consequently the related dairy products. Also, monitoring programs should be more extensive with a particular attention in monitoring aflatoxin in milk and milk products. The milk and milk products should be kept under refrigeration and the practice of display at room temperature should be discouraged.

 

REFERENCES

 

A.P.H.A. “American Public Health Association“ (1992): Standard methods for the examination of dairy produts 16th Ed., American Public Health Association. Washington.

Al-Hawary, I.I.; Sobeih, A.M.K. and Aman, I. (2002): Microbiological evaluation of cheeses and milk powder in Kafr El-Sheikh and El-Gharbia governorates. Minufyia Vet. Med. J. 2 (1): 265-276.

Amer, A.A.A. (2002): Safety and quality of butter and cheese through mycological criteria Thesis, Ph.D., Fac. Vet. Med., Alexandria Univ., Egypt.

Bailey, W.R. and Scott, E.G. (1978): Diagnostic microbiology. A textbook for the isolation and identification of pathogenic microorganisms. The C.V. Mosby by company, Saint Louis, Toronto, London.

Barrios, M.J.; Medina, L.M.; Cordoba, M.G. and Jordano, R. (1997): Aflatoxin producing strains of aspergillus flavus isolated from cheese. J. food Prot. 60: 192-194.

El-Asuoty, M.S.M.I. (2011): Mycological evaluation of some dairy products with special references to mycotoxins production, Thesis, Ph.D., Fac. Vet. Med., Alexandria Univ., Egypt.

El-Diasty, E.M. and El-Kaseh, R.M. (2009): Microbiological monitoring of raw milk and yoghurt samples collected from El-Beida city. Arab J. Biotech, 12, (1): 57-64.

El- Komy, K. (2002): Pollution of some street vended dairy products by moulds of public health hazard. Thesis, M.V.SC., Fac. Vet. Med., Alexandria Univ., Egypt.

El-Shrief, L.M.T. (2000): Incidence of mycoflora and some mycotoxins in locally manufactured cheese. Thesis, M.V.Sc. Fac. Vet. Med., Assiut Univ., Egypt.

Ghazal, G.H. (2001): Mycological studies of raw milk and cheese. Ph.D. Thesis, Fac. of Vet. Med., Zagazig Univ., Egypt.

Gqaleni, N.; Smith, J.E.; Lacey, J. and Gettinby, G. (1997): Effects of temperature, water activity and incubation time on production of aflatoxins and cyclopiazonic acid by an isolate of Aspergillus flavusi surface agar culture. J. Appl. Environ. Microbiol., 63: 1048-1053.

Gulbe, G. and Valdovska, A. (2014):Diversity of microscopic fungi in the raw milk from Latvian organic farms, Proc. Latv. Uni. Agr., 2014, 31 (326) 46-53.

Hafez, D.A.A. (2010): Sanitary condition of some processed milk products sold in Beni-Suef city. Thesis, M.V.SC. Fac. Vet. Med., Beni-suefuniv., Egypt.

Hassan, G.M. (2003):Quality assessment of some dairy products at consumer level. Thesis, Ph.D., Fac. Vet. Med., Cairo Univ. Beni-suef branch, Egypt.

Hup, H. and Stadhounders, R. (1972): In “Food and Beverage Mycology” 2nd ed. (Lorry R. Bevchat, 1978). AVI publishing company INC, wesport, Connecticut pp:484.

Karthikeyan, N. and Pandiyan, C. (2013): Microbial quality of Khoa and Khoa based milk sweets from different sources. Int Food Res. J. 20(3): 1443-1447.

Khair-Allah, H.M. (2000): Safety and quality of some dairy products through microbial criteria. Thesis M.V.SC., Fac. Vet. Med. Alexandria Univ.

Kure, C.F.; Skaar, I. and Brendehaug, J. (2004): Mold contamination in production of semi hard cheese Int. J. food Microbiology. 15:93:41-49.

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Lavoie, K.; Touchette, M.;  St-Gelais, D. and Labrie, S. (2012):Characterization of the fungal microflora in raw milk and specialty cheeses of the province of Quebec, Dairy Sci Technol. 2012 Sep; 92(5): 455–468.

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Martins, M.; Martins, H.; Lowdes, M. and Lourdes, H. (1995): Occurrence of aflatoxin, ochratoxin, steromgasten and patulin in portuguess cheese, veterinariatechnia, 5(1):18.

Moawad, A.A.; Galal, E.A.; Abd El-Hady, H.M. and Dardir, H.A. (2002): Role of dairy plant in improving some aspects of kareish cheese. Egypt. Vet. Med., Ass., 62, (2): 157-165.

Moustafa, M.F. (2004): Studies on Escherichia coli O157: H7 in milk and some dairy products. Thesis, Ph.D, Fac. Vet. Med., Assiut Univ., Egypt.

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Pešić-Mikulec, D.; Stojanović, L. and Jovanović, L. (2005): Moulds associated with milk depending on macroclimate and geographical location. App Ecol Environ Res 3(2): 61-65.

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Sengun, I.; Yaman, D. and Gonul, S. (2008): Mycotoxins and mould contamination in cheese World Mycotox. J., 3: 291-298.

Škrinjar, M.; Jakić-Dimić, D.; Blagojev, N. and Šošo, V. (2011): Results of one-year investigations of the contamination of dairy cattle feed and raw milk with moulds and mycotoxins. Biotechnol Anim Husb, 27(3): 985-992.

 

 

 

التقييم الميکولوجى للحليب وبعض منتجات الالبان فى مدينه بنى سويف

 

سعديه حلمى الشناوى ، عادل مصطفى الخولى ، عرفه مشرف سليمان مشرف ، سمر وحيد شرقاوى

 

E-mail: ashrykhaled@gmail.com      Assiut University web-site: www.aun.edu.eg

 

تم إخضاع 125 عينة من الحليب الخام ، الجبنة القريش المصنعة محليا ، الجبن الصلب (الجبنة الرومى) ، الزبادي العادي والزبادى المنکهة (25 من کل منها) التي تم جمعها من محلات الألبان والأسواق في مدينة بني سويف لفحوص فطرية. تم الکشف عن الخمائر والعفن في32٪ و36.8٪ من مجموع العينات التي تم فحصها على التوالي. تم الکشف عن الخمائر في الحليب وعينات الجبن القريش المصنعة محليا والجبنة الرومى بقيمة متوسطة 6.22Χ102±3.62Χ102 و3.23  Χ106±2.017Χ106 و 17±7.21 على التوالي، في حين أن الزبادي العادي والزبادي المنکهة کان خالي من أي خمائر. وکانت الاصناف الشائعه الأکثر عزلا الکانديدا سيدو تروبيکالز ، الرودوتورولا، اصناف من الکانديداو التوريولوبسيس (کانديدا) جلابراتا وکربتوکوکس نيوفورمانز. وکان متوسط ​​قيمة العفن 2.23Χ 102 ±9.3Χ101 و 1.068Χ106±7.20Χ105 و 4.8±1.54 و 2.4±0.871 في الحليب المفحوص ، الجبنه القريش المصنعة محليا ، الجبن الرومى والزبادي المنکه على التوالي ، في حين لم يتم الکشف عن العفن في فحص عينات الزبادي العادي. وکانت الاصناف الشائعه الأکثر عزلا الميوکور، الأسبريجيلوس فلافوس، الأسبريجيلوس نيجر، الأسبريجيلوس فيوميجاتس، الأبسيديا، البينيسيليوم، الجيوتراکوم کانديدم، الکلادوسبوريوم ، الفوما والفيوزاريوم. وکانت جميع عينات الجبن الرومى التي تم فحصها خالية من الأفلاتوکسين M1 معايرة من قبل جهاز الفلورومتر. وقد تمت الاشاره إلى الاهمية الاقتصادية والصحة العامة لوجود الفطريات والسموم الفطرية وکذلک الاحتياطات الصحية.

 

 

REFERENCES
 
A.P.H.A. “American Public Health Association“ (1992): Standard methods for the examination of dairy produts 16th Ed., American Public Health Association. Washington.
Al-Hawary, I.I.; Sobeih, A.M.K. and Aman, I. (2002): Microbiological evaluation of cheeses and milk powder in Kafr El-Sheikh and El-Gharbia governorates. Minufyia Vet. Med. J. 2 (1): 265-276.
Amer, A.A.A. (2002): Safety and quality of butter and cheese through mycological criteria Thesis, Ph.D., Fac. Vet. Med., Alexandria Univ., Egypt.
Bailey, W.R. and Scott, E.G. (1978): Diagnostic microbiology. A textbook for the isolation and identification of pathogenic microorganisms. The C.V. Mosby by company, Saint Louis, Toronto, London.
Barrios, M.J.; Medina, L.M.; Cordoba, M.G. and Jordano, R. (1997): Aflatoxin producing strains of aspergillus flavus isolated from cheese. J. food Prot. 60: 192-194.
El-Asuoty, M.S.M.I. (2011): Mycological evaluation of some dairy products with special references to mycotoxins production, Thesis, Ph.D., Fac. Vet. Med., Alexandria Univ., Egypt.
El-Diasty, E.M. and El-Kaseh, R.M. (2009): Microbiological monitoring of raw milk and yoghurt samples collected from El-Beida city. Arab J. Biotech, 12, (1): 57-64.
El- Komy, K. (2002): Pollution of some street vended dairy products by moulds of public health hazard. Thesis, M.V.SC., Fac. Vet. Med., Alexandria Univ., Egypt.
El-Shrief, L.M.T. (2000): Incidence of mycoflora and some mycotoxins in locally manufactured cheese. Thesis, M.V.Sc. Fac. Vet. Med., Assiut Univ., Egypt.
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