(summary of ph.d. thesis) · highly productive breed: alpina francez, saanen -ul elvecian, nobila...

UNIVERSITY OF AGRICULTURAL SCIENCE ANDVETERINARY MEDICINE CLUJ-NAPOCA

DOCTORAL SCHOOLFACULTY OF AGRICULTURE

bio. chim. ZORICA MARCELA P. VOŞGAN

MICROBIOLOGICAL RESEARCH ON SOIL - PLANT – GOAT MILK

TRACEABILITY UNDER THE ECOPEDOLOGICAL CONDITIONS OF

ŞURDEŞTI VILLAGE, MARAMUREŞ COUNTY

(SUMMARY OF Ph.D. THESIS)

SCIENTIFIC COORDINATOR :

Ph.D. Univ. Prof. ROXANA VIDICAN

CLUJ-NAPOCA

2013

2

TO,

Mr. (Mrs.) ---------------------------------------------------------------

Please you receive a copy of the summary of the thesis entitled: ,, MICROBIOLOGICAL

RESEARCH ON SOIL - PLANT – GOAT MILK TRACEABILITY UNDER THE

ECOPEDOLOGICAL CONDITIONS OF ŞURDEŞTI VILLAGE, MARAMUREŞ COUNTY”

edited by bio. chim. Zorica Marcela VOŞGAN, to obtain scientific title ,, Doctor in Agronomy “

Publishes support of the PhD thesis will take place on 02.10.2013, at 10.00, Green

Amphitheatre, of USAMV Cluj-Napoca.

Commission of PhD has the following structure:

PRESIDENT: Conf.univ. dr. Dan VÂRBAN

- Faculty of Agriculture –USAMV Cluj-Napoca

SCIENTIFIC COORDINATOR: Ph.D. Univ. Prof. Roxana VIDICAN

- Faculty of Agriculture –USAMV Cluj-Napoca

OFFICIAL REVIEWERS:

Ph.D. Univ. Prof. Ioan ROTAR-Faculty of Agriculture-USAMV Cluj-Napoca

Ph.D. Univ. Prof. Neculai DRAGOMIR – USAMVB Timişoara

Ph.D. Univ. Prof. Luminiţa COJOCARIU – USAMVB Timişoara

Assessments, comments and suggestions, please send them to the address USAMV,

ClujNapoca, Mănăştur Way, no. 3-5, code 400372.

The thesis is submitted to the Library of USAMV Cluj-Napoca, where it can be found.

drd. Zorica Marcela VOŞGAN

Scientific coordinator: Ph.D. Univ. Prof. ROXANA VIDICAN

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CONTENT

INTRODUCTION........................................................................................................... 5

CHAPTER I FOOD TRACEABILITY.......................................................................... 6

CHAPTER II QUALITY TRAITS OF GOAT MILK.................................................. 7

2.1. Importance and evolution of goat growth.......................................... 7

2.2. Properties of goat milk....................................................................... 7

2.3. Microbiological activity in goat milk-specific and non-specific

microbiota.......................................................................................... 8

CHAPTER III RESEARCH OBJECTIVES AND THE NATURAL SETTING.......... 10

3.1. The purpose and objectives of the PhD thesis................................... 10

3.2. Geographical, geomorphologic and climatic characterization of the

tested area.......................................................................................... 11

3.2.1. Locating research………………………………................ 11

3.2.2. The climate in the area investigated.................................... 11

CHAPTER IV MATERIAL AND METHOD.............................................................. 13

4.1. Analysis of physical-chemical characteristics of the soil in the

investigated area................................................................................ 13

4.2. Analysis of grassland vegetation ...................................................... 14

4.3. Analysis of physical, chemical and microbiological characteristics

of goat milk........................................................................................ 15

CHAPTER V RESULTS AND DISCUSSION............................................................ 17

5.1. Şurdeşti village soil analysis.............................................................. 17

5.2. Gutâi Mountains soil analysis ........................................................... 18

5.3. Analysis of vegetation in the village Şurdeşti................................... 18

5.4. Analysis of vegetation on Gutâi Mountains pastures........................ 19

5.5. Quality indicators of goat milk.......................................................... 20

5.5.1. Physical-chemical analysis of goat milk............................. 20

5.5.2. Microbiological analysis of goat milk................................ 20

5.5.2.1. Total number of aerobic mesophilic germs....... 21

5.5.2.2. Total number of yeasts and molds..................... 21

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5.5.2.3. Number of coliform bacteria............................. 21

5.5.2.4. The number of colony forming units of

Escherichia coli β-glucuronidase-positive........ 22

5.5.2.5. The number of coagulase-positive

staphylococci (Staphylococcus aureus and

other species)..................................................... 22

5.5.2.6. Bacteria of the genus Salmonella...................... 22

5.5.2.7. The number of somatic cells............................. 22

5.6. Traceability soil-plant-animal origin product.................................... 23

CHAPTER VI CONCLUSIONS AND RECOMMENDATIONS.............................. 29

6.1. Conclusions...................................................................................... 29

6.2. Recommendations............................................................................ 30

SELECTIVE BIBLIOGRAPHY................................................................................... 31

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INTRODUCTION

Consuming animal products, which occupies an important place in the rational

diet of man, is not without danger. Therefore special attention should be paid to the type

of feeding, supervising the product at its source.

Special measures on food safety, ensuring the traceability of food from its source

to the consumer, have been taken in the last years. The traceability systems allow the

notification and correction or the removal of the risk factors, for making quality products.

Of all ruminants that are grown in our country, the goat was placed on one of the

last places of interest within the livestock, although it has the ability to provide high

quality food.

Goat milk is the natural food that contains all the nutrients needed for growth and

body development.

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CHAPTER I

FOOD TRACEABILITY

The concept of traceability has become well-known in most of the food industry,

the food market becoming extremely dynamic. This trend is explained by the increased

exigencies and the diversity of the customers’ requirements, so the manufacturers are

challenged to develop and acquire new products to meet all aspects of consumer safety

and quality requirements (STĂNCIUC and ROTARU, 2009). According to ISO

22005:2007, "traceability is the ability to trace the history, application or location of an

item via the recorded information”.

Traceability is a concept which is valid for all food products (STĂNCIUC and

ROTARU, 2009); its importance considerably increased, much more in the last decade

(MUNTEAN and RADU, 2007), but although it is believed to be a simple concept, is

complex in the terms of implementation.

Firstly, traceability can be used as a management instrument of the risks linked to

the food safety and the issues regarding animal safety, and secondly, it has an important

role in establishing credibility on the market. All consumers have the right for good

quality food. FAO has been a frontrunner to upgrade the member states capacity, to

establish and implement food safety and adequate control systems of the food quality.

For the agro-industrial sector of this country a strategy has been elaborated, which

is to consider the market exigencies as well as the potential risk factors, because

agriculture is not only a source of agricultural raw materials, but also of exploiting food

products to ensure public health (BALŞ, 2010).

Due to their perishability, fresh foodstuff requires traceability. In case of milk and

dairy, traceability is very important in testing the product authenticity and origin. The

quality of the final product of the entire dairy chain starts with the diet of animals, the

species of plants which are present in fodder (PONZONI et al., 2009).

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CHAPTER II

QUALITY TRAITS OF GOAT MILK

2.1. IMPORTANCE AND EVOLUTION OF GOAT GROWTH

According to TAFTĂ, 2002, within the goat raising countries, India and China are

situated on the first place, followed in decreasing order by Pakistan, Nigeria, Somalia,

Sudan, Brazil, Indonesia, Mexico, etc. Goat rearing is also well developed in some

European countries: France, Swiss, Austria, Great Britain, Portugal etc., due to some

highly productive breed: Alpina franceză, Saanen-ul elveţian, Nobila germană.

In this country, the perspective of the goat rearing development is increasing, but

it still suffers due to the lack of an organized program of breeding and improvement.

Goats are mostly prevalent in the hilly, mountainous areas, around some towns, both on

small farms of two-three heads as well as in larger groups of 40-60 heads, even within

specialized farms (TAFTĂ, 2002). According to TAFTĂ, (2002), 70 % are goats of the

Carpatină breed, unimproved, hardy, resistant to harsh living conditions and with

relatively low milk yield, about 30 % are of the Albă de Banat breed, ameliorated, with

double milk yield.

According to the general agricultural census of 2010, the total goat stock in

Romania was of 1.240.858 heads, of which in the Maramureş County, the number of

goats was of 17.744 heads.

According to BANU et al., (2007), goats are important in economy for: milk yield,

meat obtained from goat kids, young fattening, adult goats, leather (hair, undercoat, goat

wool); leather and fur, manure that is very good for fertilizing.

2.2 . PROPERTIES OF GOAT MILK

Goat milk has a white color (lower in pigment) towards yellowish due to the

pronounced fineness of fat particles uniformly disseminated within its mass, with specific

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smell and taste that are pleasant in case of hygienic maintenance and milking and proper

feeding.

The chemical contents of goat milk is different from that of cow milk by its higher

contents of protein, which comprises of 75-80 % casein, a feature that is highly useful in

its transformation into cheese. The contents of fat and protein in goat milk is a little bit

higher than that of cow milk, the other components being relatively equal, excepting

sheep milk (TAFTĂ, 2002). Due to the fineness of fat globules, goat milk is easily

absorbed by the intestinal villi; under the action of gastric juice protein coagulates in fine

flocculants which are easier assimilated by the stomach, the same as glucose and lactose,

which gives it a higher nutritional value (TAFTĂ, 2002).

Sugars in goat milk are represented by lactose; there are small amounts of glucose,

galactose, and some oligosaccharides. The lactose content in goat milk is about 0.2-0.5%

lower than in cow milk (POSATI and ORR, 1976; HAENLEIN and CACCESE, 1984;

CHANDAN et al., 1992).

Goat milk is rich in A vitamin, due to the remarkable ability of goats to convert

carotene pigments. Vitamin A is found in its final form in goat milk, whereas in cow milk

it is in the form of carotene that is why goat cheese and butter are white (SPÄTH and

THUME, 2008).

Goat milk is richer in calcium, phosphorus, citric acid, potassium, magnesium, and

having a high content of vitamin A it is indicated in the nutrition of infants and the

elderly (TIŢA, 2002).

2.3. MICROBIOLOGICAL ACTIVITY IN GOAT MILK-SPECIFIC AND

NON-SPECIFIC MICROBIOTA

Contamination of milk could be made from various sources, and microbial load

could be quantitatively and qualitatively quite varied, depending on the conditions under

which milk is produced, handled and processed. Microorganisms reach food from natural

or external sources as a result of processing and handling, until the moment of

consumption. (VANDERZANT and SPHITTSTOESSER, 1992; AONOFRIESEI, 2012).

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Milk microbiota generally consist of pathogenic microorganisms, as well as

facultative pathogenic or pathogenic ones.

The category of non-pathogenic microorganisms includes lactic bacteria, largely

distributed in nature, which play an important role in the fermentation of food and feed.

By the milk contamination with scavenge germs, the risk of including pathogen germs

increases. Many pathogenic bacteria do not multiply in milk (Mycobacterium

tuberculosis, M. bovis, Brucella, Rickettsia), their danger depending on the initial degree

of milk contamination, the dilution of subsequent treatments, the elapsed time until the

consumption of milk and other factors. Other pathogen germs could multiply in milk

(Staphylococcus aureus, Streptococcus pyogenes, Escherichia coli, etc.).

The metabolic activity of most pathogens is inhibited at low temperatures, so it is

very important to immediately cool the milk after milking, until its heat treatment.

The milk contamination with yeast and mold is inevitable, especially in case of

manual milking. Species of the genera: Torula, Candida, Saccharomyces, Mucor,

Aspergillus, Penicillium, Cladosporium, Oidium, etc. could be responsible for modifying

the organoleptic properties (BĂRZOI and APOSTU, 2002).

Also, milk somatic cells are part of the hygienic quality indices of milk for human

consumption.

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CHAPTER III

RESEARCH OBJECTIVES AND THE NATURAL SETTING

3.1. THE PURPOSE AND OBJECTIVES OF THE PhD THESIS

The aim of the thesis is to determine the traceability of goat milk, studying the

whole chain: soil-plant-animal product, correlations that are between these links and that

determine the quality of the final product.

To achieve the overall objective of the thesis we propose the following specific

objectives:

1. Pedological characterization of the studied perimeter, meaning Şurdeşti village

pasture respectively Şurdeşti village common pasture from Gutâi Mountain:

the study of soil morphology: the sequence of genetic horizons, color and structure

of the horizons;

physical-chemical characterization of the soil from the tested field: texture, the

hygroscopic coefficient, pH, humus, soil nutrients (nitrogen, phosphorus and

potassium), cation adsorption capacity;

assessment of the content of microelements in soil;

2. Characterization of grassland vegetation from Gutâi Mountain Area where

livestock grazing takes place:

vegetation qualitative assessment for calculating the pastoral value of the

meadows;

determining the production capacity of grasslands in order to establish the

organization of grazing;

3. Analysis of the quality characteristics of goat milk, considering both

physicochemical and microbiological indicators:

determining the quality of goat milk produced by a traditional production system,

depending on the season, respectively goats grazing area;

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study of the specific saprophytic milk microflora (total aerobic mesophilic germs/

ml, total yeast and molds/ml) and also of the facultative pathogen or pathogen

agents (coagulase-positive staphylococci, coliforms, Escherichia coli, Salmonella

sp.);

4. Establishing the correlation between soil-plant-animal product (goat’s milk) in

the area where goats graze (South West Region of the Gutâi Mountains) and observing

the role of traceability on animal origin product.

3.2 . GEOGRAPHICAL, GEOMORPHOLOGIC AND CLIMATIC

CHARACTERIZATION OF THE TESTED AREA

3.2.1. Locating research

The research takes place under ecopedological conditions in the Gutâi Mountains

perimeter (Maramureş County, Northwestern Romania) in two geographical points:

Şurdeşti village pasture, a hilly area situated at an altitude of 600-900 m (in the

vicinity of relief units Bulbucul Mic, Bulbucul Mare, Piatra Roşie, Muta Mare, Muta

Mică, Dealul Crucii, Arşiţa, etc.), occupying an area of about 70 hectares.

Gutâi Mountains pasture (the Southwestern part of the massif) situated at an

altitude of 1000-1300 m within an area of about 133 ha (according to Şişeşti Village

Hall), (Figure 1, 2).

3.2.2. The climate in the area investigated

The experimental area is characterized by a temperate continental climate with late

springs and sufficient rainfall throughout the year, with an average annual temperature of

9.4°C, also the average temperature of the hottest month is 39.2°C and of the coldest

month is -30°C.

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Fig. 1. Location of the Sisesti village in the Maramures County

Fig. 2. Location of the research place (http://www.google.com/mapmaker?hl=ro)

A- Common pasture of Şurdeşti village from Gutai MountainB- Şurdeşti village pasture

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CHAPTER IV

MATERIAL AND METHOD

To achieve the objectives proposed in the thesis, the experience took place over

three years respectively between 2010-2012, in the Gutâi Mountains Area (Maramureş

County), on goat milk originated from a herd of 50-60 goats bred and maintained

extensively.

In order to accomplish the objectives considering the traceability of goat milk,

starting from the genetic type of soil - vegetation type - up to the microbiological

characteristics, there were performed the following analyzes:

Analysis of physical-chemical characteristics of the soil cover from the

experimental area;

Analysis of experimental field vegetation;

Analysis of quality characteristics (physical, chemical and microbiological) of

goat milk;

4.1. ANALYSIS OF PHYSICAL-CHEMICAL CHARACTERISTICS OF

THE SOIL IN THE INVESTIGATED AREA

Physical and chemical analyses of the soil in the two experimental points were

performed at O.S.P.A. Cluj-Napoca and in the laboratories of the University of

Agricultural Sciences and Veterinary Medicine Cluj-Napoca.

Were collected:

12 soil samples from three points analyzed on Şurdeşti village pasture;

17 soil samples from three points analyzed on Gutâi Mountains pasture;

Working methods applied to characterize the physico-chemical soil type are

presented in Figure 3.

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Fig.3. Methods of soil analysis

4.2. ANALYSIS OF GRASSLAND VEGETATION

To determine the type of grassland in the study area, we conducted a recognition

travel itinerary and noted the species encountered after an assessment scale developed by

Braun-Blanquet (1928, 1951).

The analysis of the floristic composition was performed by the geobotanical or

phito-social methods (ROTAR et al., 2009).

Determination of grassland production – an usual method for the assessment of

the green mass production in our country is a gravimetric method consisting of mowing

and weighing the green mass of a certain number of evidence wedges (based on meadows

uniformity and analyzed surface) before each harvest cycle. Results are extrapolated to

the unit area (ha) (ROTAR et al., 2009).

1. Taken from Şurdeşti: threesampling points.

2. Taken from Gutai massif:three sampling points.

(ICPA within the O.S.P.A.Maramureş)

1. Particle size analysis - pipette method ofinterpreting the results after ICPA -Kacinski, (ICPA, 1981);

2. The hygroscopic coefficient, STAS7184/6-78;

3. Soil reaction (pH-ul), STAS 7184/13-88;4. Determination of the humus content (%),

STAS 7184/21-82;5. Determination of total nitrogen content

(N-total %) - Kjeldahl method;6. Determination of potassium by the flame-

photometric method, ICPA, 1981;7. Colorimetric determination of

phosphorus, ICPA, 1981;8. Exchangeable hydrogen (SH-me/100g

ground) Cernescu by percolation method,ICPA, 1981;

9. The amount of basic cations (SB-me/100gground)- Kappen method, ICPA, 1981;

10. Trace elements in soil-AAS method;

Soil samples

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Vegetation qualitative assessment and determination of the pastoral value -

Pastoral value (Vp) is a synthetic indicator which includes the main elements of floristic

composition, cover percentage of each species and the field value of the included species

(ROTAR et al., 2009).

4.3. ANALYSIS OF PHYSICAL, CHEMICAL AND MICROBIOLOGICAL

CHARACTERISTICS OF GOAT MILK

Milk samples were collected by hand milking of 50-60 goats grown and

maintained in the traditionally system (Figure 4), the predominant race being the

Carpatină one, rather than the Albă de Banat.

For the physical-chemical analysis there were collected 10 samples of goat milk,

depending on the grazing time of the goats: spring on Şurdeşti village pasture, summer on

Gutai Mountains pasture and in fall again on the Şurdeşti village pasture.

Microbiological analysis was also carried out according to the grazing period,

three milk samples being collected in each season.

The milk samples were analyzed in specialized laboratories:

Teaching and research laboratories of the Department of Chemistry-Biology,

Faculty of Science, in the North University Center of Baia Mare, Technical University of

Cluj Napoca.

The laboratories of the Sanitary Veterinary and Food Safety Directorate

(SVFSD) Maramureş.

Fig. 4. Aspects of goat milk production (original)

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Physical-chemical methods and microbiological analysis of goat milk are

presented in Figure 5.

Fig. 5. Methods of analysis of goat milk

Goat milk samples

Sampling of milk by handmilking

STAS 6349/1-80

Physico-chemical indicators Microbiological indicators

1. Acidity °T - titration method;

2. Density or specific gravity of milk -

with termolactodensimetru;

3. Dry matter (D.M.)- drying oven

method (1020C -1050C) ;

4. Determination of total protein

Kjeldahl method;

5. Determination of fat content -

Method acid-butirometrica;

6. Determining the amount of lactose

in milk - Potassium Ferricyanide

Not method;

7. Determination of total ash - STAS

6357-75;

(POP, 2008)

1. Total number of aerobic mesophilic

germs (TNG/ml; CFU/ml), STAS

ISO 4833-2003;

2. Total number yeasts and molds, STAS

ISO 7954-2001;

3. The presence and number of coliform

bacteria, STAS ISO 4831/1992;

4. The number of colony forming units of

Escherichia coli β-glucuronidase-positive,

STAS ISO 16649-2/2007;

5. The number of coagulase-positive

staphylococci (Staphylococcus aureus and

other species), STAS ISO 6888-1/2002;

6. Bacteria of the genus Salmonella, STAS

ISO 6579:2002;

7. The number of somatic cells, APOSTU

şi ROTAR, 2009;

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CHAPTER V

RESULTS AND DISCUSSION

5.1. ŞURDEŞTI VILLAGE SOIL ANALYSIS

In Şurdeşti village there were determined the physical-chemical properties of soil

in three points, in areas where goats graze frequently during spring and autumn. As a

result of the performed analysis, according to the Romanian System of Soil Taxonomy

(SRTS) in the studied area there were identified the following genetic types of soil:

Luvosol located at 718 m altitude, is characterized by El eluvial horizon and B

argic horizon (Bt) specific to luvosoils. It is characterized by a moderately acidic pH with

values between 4.6-5.1, a reduced supply in essential elements, the degree of base

saturation is 41-60 %, cation exchange capacity is low (T <17.4 me/100g soil).

Districambosoil gleic located at an altitude of 746 m, presents the following

sequence of genetic horizons: Ao-A/B-Bv-CGr. Acidity is pronounced in the superficial

horizons, tending to neutralize at depth over 80 cm, humus content is decreased, only

3.12 % in the Ao horizon, so that the total nitrogen supply is moderate at the surface to

absent in the depth. The degree of base saturation is below 70 % and cation exchange

capacity is low.

Regosol located at 731 m altitude, is a pH acidic soil, poor in humus and nutrients.

The degree of base saturation is below 70 % and cation exchange capacity is at most

12.68 me/100 g soil.

Content analysis results for soil microelements of Şurdeşti village show an excess

of the normal value for Pb, being surpassed both the alert threshold and threshold for

intervention, while other micronutrients, Cu, Zn, Mn, Ni, Cd, do not exceed the normal

values for the three soil types identified.

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5.2. GUTÂI MOUNTAINS SOIL ANALYSIS

Following the researches on the South-western Region of Gutâi Mountain there

was identified one genetic type of soil in all three points where analyzes were performed:

districambosoil typical.

This type of soil is characterized by a strong acid reaction, high humus content in

the upper layers, since due to the pronounced acidity and cool environment,

decomposition and mineralization is very slow, as indicated by the degree of base

saturation of the soil which is <70 %. Nutrient content is low and the concentration of

trace elements such as Pb, Cd, Zn, is beyond the normal limit, however Cu, Mn, Ni, do

not exceed the permissible values.

5.3. ANALYSIS OF VEGETATION IN THE VILLAGE ŞURDEŞTI

On Şurdeşti village pasture with an area of 70 ha, there were found the following

plant associations:

1. Festuco rubrae – Agrostietum capillaris, Horvat 1951

2. Violo declinatae-Nardetum, Simon 1966 (Syn. Nardetum strictae

montanum, Resmeriţă et Csűrös 1963; Nardetum strictae subalpinum, Buia et al. 1962;

Nardetum strictae alpinum, Puşcaru et al. 1956; Nardetum alpigenum austro-carpaticum,

Borza 1959; Nardo-Vaccinietum, Resmeriţa 1970).

On the studied grassland, the hemicryptophytes are of numeric majority, while the

Eurasian species stands in the geoelements category. Taking into account the types of

plants, depending on the main environmental indices, the majority is mesophilic species,

based on the heat index, dominant species are micro-mesothermal, as considering the

chemical reaction of the soil, the euriionic species are predominant. In terms of cariology,

diploids are in greater number and the economic value of grassland is a medium one.

The meadow has an averaged pastoral value of 1.9, with a score of 41-50 points of

evaluation, it is the sixth category, medium, supporting a load of 0.81 to 1.00 LU/ha. The

production of green mass is 13.37 t/ha, refusals R = 1.51 t/ha, edible production is 11.86

t/ha, and the coefficient of grass utilization is K = 88.7%.

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5.4. ANALYSIS OF VEGETATION ON GUTÂI MOUNTAINS PASTURES

The investigated perimeter of Gutâi Mountains (Gol Gutin and Valea Docăi)

occupies an area of 133 ha; wooded meadows alternate with meadows on which the

following plant associations are predominating:

1. Violo declinatae-Nardetum, Simon 1966 (Syn. Nardetum strictae montanum,

Resmeriţă et Csűrös 1963; Nardetum strictae subalpinum, Buia et al. 1962; Nardetum

strictae alpinum, Puşcaru et al. 1956; Nardetum alpigenum austro-carpaticum, Borza

1959; Nardo-Vaccinietum, Resmeriţa 1970)

2. Agrostietum stoloniferae, (Ujvárosi 1941) Burduja et al. 1956 (Syn. Rorippo

syilvestris- Agrostietum stoloniferae, (Morr 1958) Oberdorfer et Th. Müller 1961)

3. Scirpetum sylvatici, Ralski 1931, Maloch 1935 em. Schwick 1944

4. Calthetum laetae, Krajina 1933

5. Campanulo abientinae-Vaccinietum myrtilli, (Buia et al. 1962) Boşcaiu 1971

(Syn. Vaccinietum myrtilli, Buia et al. 1962; Juncetum trifidi- Vaccinietum myrtilli,

Resmeriţă 1976)

In the studied grasslands, the bioforms statistics show that the hemicryptophytes

have the largest share, while of geoelements, the dominant species are Eurasian.

Ecological analysis reveals a dominance of the mesophilic species, depending on the

thermal factor amphitolerant and micro-mesothermal species are predominant, the soil

reaction indicates the dominance of euriionic species, alongside which many plants are

acidophilic and acid-neutrophil ones. In these associations’ phytocoenosis, polyploids are

ahead of the diploids number, and the economic value of forage is low.

Meadow has a pastoral value of 0.85, with a score of 20 points of evaluation, it is

category IX, degraded, carrying a load of 0.21 to 0.40 LU/ha. The production of green

mass is 11.6 t/ha, refusals R = 6.15 t/ha, edible production is 5.45 t/ha, and the coefficient

of grass utilization is K = 46.9%.

20

5.5. QUALITY INDICATORS OF GOAT MILK

5.5.1. Physical-chemical analysis of goat milk

Considering the same species and breed, feeding mode and lactation period, has a

great influence on the composition of milk.

Physical-chemical parameters of goat milk based on grazing area (13.705% DM

0.8449% ash, 4.318% protein, fat 4.203%, 4.081% lactose, spring, 12.3% DM, 0.7881%

ash, 3.46% protein, fat 4.003%, 4.163% lactose, summer, or 16.84% DM, 0.95% ash,

5.381% protein, 7.00% fat, 4.41% lactose, fall) show a decrease in the quality of milk

during summer when goats graze in Gutâi Mountains.

Variability of physical-chemical parameters of goat milk collected by manual

milking in pen, depending on the season, but also on the grazing territory is presented in

Figure 6.

0123456789

1011121314151617

Spring Summer Autumn

Season

The

ave

rage

s val

ues o

f phy

sico-

chem

ical

para

met

ers

Density, g/cm3

Dry substance, %

Ash, %

Protein, %

Fat, %

Lactose, %

Fig. 6. The physical-chemical variability and average values of goat milk

depending on the season

5.5.2. Microbiological analysis of goat milk

Aseptically collected milk samples were microbiological analyzed during three

seasons (spring, summer, autumn), by the cessation of the lactation. Meanwhile milking

21

was done by hand; respecting hygiene as possible considering the animal (goat udder and

body hygiene), the milker and milking environment.

5.5.2.1. Total number of aerobic mesophilic germs (TNG/ml or CFU/ml)

In spring, when the quantity of milk is larger and the feeding consists of raw

vegetation, total aerobic mesophilic germs (TNG) is of 3108.77 CFU/ml, a small

number compared to the number of microorganisms determined during summer

( 3103.97 CFU/ml) , when the vegetation is poor, or during fall ( 3105.81 CFU/ml)

characterized by a low milk production.

Even if the microbial load in goat milk is higher in summer, when maintenance

conditions are more difficult, TNG is not exceeding 100.000/ml milk according to

European regulations.

Microscopic examination of the native preparations and smears indicates the

presence of species of the genus: Lactobacillus sp., Pediococcus sp., Streptococcus sp.,

Lactococcus sp.

5.5.2.2 Total number yeasts and molds

The average values obtained for the total number of yeasts and molds (TNYM)/ml

goat milk in the three seasons are: 0.8 × 102 CFU/ml yeasts, 0.3 × 102 CFU/ml molds

(spring), 26, 8 × 102 CFU/ml yeasts, 10.6 × 102 CFU/ml molds (summer) and 2.9 × 102

CFU/ml yeasts, 0.7 × 102 CFU/ml molds (autumn).

By microscopic examination have been found yeast such as Rhodotorula sp.,

Cryptococcus sp. (only in the samples of milk analyzed in summer), Kluyveromyces sp.

and molds: Cladosporium sp. Fusarium sp. Penicillium sp. Mucor sp.

5.5.2.3. Number of coliform bacteria

Coliforms are indicators of fecal contamination of livestock products and their

presence was highlighted by the gas bubbles which appeared in the Durham fermentation

tubes after incubation at 370C.

The results show the presence of coliforms in goats’ milk: 40 coliforms/ml

(spring), 150 coliforms/ml (summer), 110 coliforms/ml (autumn).

22

5.5.2.4. The number of colony forming units of

Escherichia coli β-glucuronidase-positive

In the coliform group of food microbiology, Escherichia coli species is

representative and it was assigned the role of opportunistic pathogen with high

circulation in nature (Barzoi and APOSTU, 2002). The presence of pathogenic

microorganisms in milk can occur either through fecal contamination or by direct

excretion from the udder in milk (MUEHLHERR et al., 2003).

Determination of the number of E. coli - glucuronidase - positive from goat milk

samples showed absence of this strain in the samples analyzed in spring and autumn, but

her presence was detected in the samples analyzed in summer, with an average of 66.7

CFU/ml milk goat.

5.5.2.5. The number of coagulase-positive staphylococci(Staphylococcus aureus and other species)

Seasonal variation of the mean number of coagulase-positive staphylococci is as

follows: 3104.1 CFU/ml, 3102.3 CFU/ml, 3108.2 CFU/ml, spring, summer and fall.

In the studied case, contamination could occur during unhygienic milking, when

pathogen transmission is effected by contaminated hands, and not only, as say KOUSTA

et al., (2010), possible contamination with S. aureus (for example) may occur in raw milk

from infected mammary glands.

5.5.2.6 Bacteria of the genus Salmonella

In all three seasons when the goat milk samples were analyzed, Salmonella sp.

was not identified in 25 ml milk, which shows the correspondence to the microbiological

standards according to Order M.H. 975-1998.

5.5.2.7 The number of somatic cells

The microbiological quality of milk depends heavily on the health of the animal's

udder, and for a good assessment there has to be determined the number of somatic cells

per ml of milk. Under the legislation of our country, somatic cell count (SCC) must be ≤

400000/ml for raw milk and dairy products.

23

The analyzed goat milk contains 509000 cells/ml in spring, 516000 cells/ml in

summer, but in autumn the number is high, of 4000000 cells/ml.

5.5. TRACEABILITY SOIL-PLANT-ANIMAL ORIGIN PRODUCT

This study research, on the Şurdeşti village pasture respectively Gutâi Mountain

pastures, showed us that there are correlations between the biological components

analyzed.

There is an obvious correlation between the trophicity degree of soil and the

grassland vegetation of the two analyzed areas (Figure 7, 8).

0

10

20

30

40

50

60

70

80

Festuco rubrae –Agrostietum capillaris

Violo declinatae-Nardetum

Vegetal associations

Num

ber

of sp

ecie

s

0

10

20

30

40

50

60

70

80

Deg

ree

of b

ase

satu

ratio

n in

soils

(V %

)

Number of species V %

Fig. 7. The relationship between the number of species in the phytocoenosis of the

Şurdeşti pasture and the degree of base saturation in soils

24

010203040506070

Violo declinatae-Nardetum

Agrostietumstoloniferae

Campanuloabientinae-

VaccinietummyrtilliVegetal associations

Num

ber

of sp

ecie

s

010203040506070

Deg

ree

of b

ase

satu

ratio

n in

soils

(V %

)

V % Number of species

Fig. 8. The relationship between the number of species in the phytocoenosis of the

Gutâi Mountains pasture and the degree of base saturation in soils

Raised acidity increases the solubility of micro-nutrients in the soil, so they can

become toxic to the plants as they are in amounts exceeding the normal values. The

acidic conditions of the soil may often increase the solubility of heavy metals such as Cu,

Zn, Pb, Mn, etc. (REDDY et al., 1995). This is observed for both Şurdeşti village as well

as Gutâi Mountains analyzed soils, (Figures 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20).

020406080

100120140160180200220240260

Ao AE El Bt Ao A/B Bt CGr Ao AC1 AC2 C

Pb m

g/kg

dry

mat

ter

0

1

2

3

4

5

6

7

8

pH o

f the

soil

Pb

PhPoint I Point II Point III

Fig. 9. The relationship between the

content of Pb (mg/kg dry matter) and the

pH of the soil in the Şurdeşti village

02468

101214161820

Ao AE El BtAo

A/B BtCGr Ao

AC1AC2 C

Cu

mg/

kg d

ry m

atte

r

0

1

2

3

4

5

6

7

8

pH o

f the

soil

CuPh

Point I Point II Point III


content of Cu (mg/kg dry matter) and the


25

0102030405060708090

100110

Ao

AE El Bt

Ao

A/B B

t

CG

r

Ao

AC

1

AC

2 C

Zn

mg/

kg d

ry m

atte

r

012345678

pH o

f the

soil

ZnPhPoint I Point II Point III


content of Zn (mg/kg dry matter) and the


0

100

200

300

400

500

600

700

800

900

Ao AE El Bt Ao A/B Bt CGr Ao AC1 AC2 C

Mn

mg/

kg d

ry m

atte

r

0

1

2

3

4

5

6

7

8

pH o

f the

soi

l

MnPh



content of Mn (mg/kg dry matter) and the


02468

101214161820

Ao AE El BtAo

A/B BtCGr Ao

AC1AC2 C

Ni m

g/kg

dry

mat

ter

012345678

pH o

f the

soil

NiPhPoint I Point II Point III

The relationship between the content of Ni

(mg/kg dry matter) and the pH of the soil

in the Şurdeşti village

0

0,2

0,4

0,6

0,8

1

1,2

Ao

AE El

Bt

Ao

A/B B

t

CG

r

Ao

AC

1

AC

2 C

Cd

mg/

kg d

ry m

atte

r 012345678

pH o

f th

e so

il

C dP hP oint I P oint II P oint III


content of Cd (mg/kg dry matter) and the


0

20

40

60

80

100

120

140

AtAo1 Ao2 A/B Bv At

AoA/B A/B Bv Cn At

AoA/B Bv1 Cn

Pb m

g/kg

dry

mat

ter

0

1

2

3

4

5

6

7

8

pH o

f the

soil

PbPh



content of Pb (mg/kg dry matter) and the pH

of the soil in the Gutâi Mountains

02468

101214161820

AtAo1Ao2A/B Bv At

AoA/BA/B Bv Cn At

AoA/BBv1 Cn

Cu

mg/

kg d

ry m

atte

r

0

1

2

3

4

5

6

pH o

f the

soil

CuPhPoint I Point II Point III


content of Cu (mg/kg dry matter) and the

pH of the soil in the Gutâi Mountains

26

0204060

80100120140

AtAo1Ao2A/B Bv At

AoA/BA/B Bv Cn At

AoA/BBv1 Cn

Zn

mg/

kg d

ry m

atte

r

012345678

pH o

f the

soil

ZnPhPoint I Point II Point III


content of Zn (mg/kg dry matter) and the pH

of the soil in the Gutai Mountains

0100200300400500600700800900

AtAo1Ao2A/B Bv At

AoA/BA/B Bv Cn At

AoA/BBv1 Cn

Mn

mg/

kg d

ry m

atte

r

012345678

pH o

f the

soil

MnPhPoint I Point II Point III


content of Mn (mg/kg dry matter) and the

pH of the soil in the Gutai Mountains

0

5

10

15

20

AtAo2 Bv At

A/B Bv AoBv1

Ni m

g/kg

dry

mat

ter

0

2

4

6

8

pH o

f the

soil

NiPh



content of Ni (mg/kg dry matter) and the pH

of the soil in the Gutâi Mountains

0

0,2

0,4

0,6

0,8

1

1,2

AtAo

1Ao

2A/

B Bv At Ao A/B

A/B Bv Cn At Ao A/B

Bv1 Cn

Cd m

g/kg

dry

mat

ter

0123

45678

pH o

f the

soil

CdPh

Point IIIPoint IIPoint I


content of Cd (mg/kg dry matter) and the

pH of the soil in the Gutâi Mountains

Converting grass in animal products allowed the extension of human living in the

high mountains above the grain areas, but unfortunately nowadays we are facing

abandonment of mountain grasslands and their poor management, which leads to a very

low level of fertilization. This is evident especially on Gutâi Mountains pasture, where

quality of goat milk is lower compared to the quality of milk when grazing on Surdesti

village pasture. The number of plant species with forage value of the identified pasture

plant associations from Şurdeşti village pasture is more numerous compared to the

number of species with forage value from Gutâi Mountains pasture.

Following the physical-chemical and microbiological parameters of goat milk,

depending on the grazing area, it was found a decreasing of milk quality when animals

27

take advantage of the pasture with low value forage species, meaning the Gutâi

Mountains pasture (Figure 21, 22).

02468

1012141618


Se ason

%

051015202530354045

N umbe r of forage spe cie s

Dry substance Ash Protein Fat Lactose N umber of forage species

Fig. 21. Variations of the physical-chemical parameters of goat milk,

depending on the fodder value of the grazing area

0

2

4

6

8

10


Season

CFU

/ml

30

32

34

36

38

40

42

Num

ber

offo

rage

spec

ies

TNG** Yeasts* Molds*Coliforms* E. coli* CPS*Salmonella sp.* Number of forage species

* CFU/ml×103

**CFU/ml×104

Fig. 22. Variation of the microbiological indicators of goat milk,

depending on the fodder value of the grazing area

Saprophytic and pathogenic microflora present seasonal variations in the analyzed

goat milk, so in spring and autumn the microbial load is lower than the average values

recorded during the summer (Figura 23).

28

00,5

11,5

22,5

33,5

44,5

55,5

66,5

77,5

88,5

99,510

TNG** Yeasts* Molds* Coliforms* E. coli* CPS* Salmonella sp.*

CFU

/ml


* CFU/ml×103

**CFU/ml×104

Fig. 23. Variation of microbiological parameters of goat milk

depending on the season

During summer milk quality decreases and this can be influenced by the

distribution of soil microelemetelor. There have been numerous studies which have

shown that high levels of micronutrients of the soil (due to pollution) may bioaccumulate

in plants. This indicates a higher risk of penetration of trace elements (Cu, Pb, Zn, Cd),

considered heavy metals, in the food chain.

Since the heavy metals exert germicidal effects, they "can quickly destroy many

types of microorganisms, including vegetative forms of bacteria, but not the endospores"

(IVANA et al., 2011). Microflora found in the analyzed goat milk varies, depending on

where the animal is grazing, but it is not influenced by the content of microelements in

soil.

Analyzing traceability of livestock products (goat’s milk) it was found that autumn

milk is superior due to its physical-chemical characteristics, nutrient content, precisely

because the number of useful microorganisms is higher in this season.

29

CHAPTER VI

CONCLUSIONS AND RECOMMENDATIONS

6.1. CONCLUSIONS

1. Analyzing the morphological and physical-chemical properties of soil in the

surveyed area (Şurdeşti locality and the south-western parts of the Gutâi Mountains,), we

have identified soils that are specific to the hilly and mountainous areas, which were

formed on substrates of igneous rocks: luvosol, gleic districambosoil, regosol and typical

districambosoil.

These are characterized by low pH, low in humus, with some exceptions in

superficial horizons, and deficient in nutrients. The contents of microelements show an

excess of the normal values for Pb; the other micronutrients, Cu, Zn, Mn, Ni, Cd, do not

exceed the normal values, with few exceptions for Zn and Cd in the soil of the Gutâi

Mountains.

2. Analysis of grassland vegetation that is grazed by goats, at Şurdeşti village

(during spring and autumn) or Gutâi (during summer), indicates a regressive trend

towards nardet.

The economic value of pasture in the Şurdeşti village is average, and that of the

Gutai Mountains is very low.

3. Analysis of the quality characteristics of goat milk produced in a semi-

subsistence farm from a flock of 50-60 heads in the Gutai Mountains indicate that the

physical-chemical and microbiological parameters of goat milk significantly varies,

depending on the season and grazing area, from the early lactation period when milking

is done three times a day to the end of the lactation, when milking is done once a day.

During spring and autumn the number of microorganisms does not exceed 82×103

TNG/ml, during summer this number increases toward the maximum value (97,3×103

TNG/ml). During summer the fungi load is higher (26,8×102 CFU/ml of yeasts, 10,6×102

CFU/ml of molds), evidently diminishing in autumn (2,9×102 CFU/ml of yeasts, 0,7×102

30

CFU/ml of molds) and spring (0,8×102 CFU/ml of yeasts, 0,3×102 CFU/ml of molds).

Also, the level of faecal contamination increases in summer (150 CFU/ml coliform) and

the colonies of E. coli β-glucuronidase positive are detected. The seasonal variation of the

number of coagulase-positive staphylococci is as follows: 1,4×103 CFU/ml spring,

3,2×103 CFU/ml in summer, or fall 2,8×103 CFU/ml, and Salmonella sp. was not

detected.

4. Between soil-plant-animal products (goat milk) obvious correlations are

established, depending on the area where goats graze, the pasture of Şurdeşti village,

respectively the pastures of the Gutâi Mountains. As such, milk yield is poor in essential

elements and has a high microbial load during summer when grazing is done in the Gutâi

Mountains.

6.2. RECOMMENDATIONS

In order to use the soil from the South Western region of Gutâi Montains in

agricultural purposes, immediate ameliorative action is needed by the application of lime

amendments.

To provide high floristic composition it is necessary to change the system of

grazing, the rotation one being indicated.

In terms of quality, the goat milk subjected to this research is good for

consumption, but requires some improvements:

Monitor the feeding of goats;

Monitoring udder health status and general health of the animal;

Monitoring of optional pathogen or pathogen microorganisms that can survive and

multiply in milk, if not processed properly and that can lead to serious

consequences for human health.

Ongoing monitoring of all key parameters of raw milk traceability allows

correlation, correction and removal of all risk factors.

It is recommended as a priority the assertion of individual farmers, small and

medium-sized farms that produce healthy food in a sustainable way.

31

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***http://www.insse.ro/cms/files/RGA2010/Rezultate%20definitive%20RGA%202010/

Volumul%20I/Tab3J-judete.pdf

***http://www.google.com/mapmaker?hl=ro

***Ordinul 975/1998 al ministrului sănătăţii privind aprobarea Normelor igienico-

sanitare pentru alimente

(summary of ph.d. thesis) · highly productive breed: alpina francez, saanen -ul elvecian, nobila...

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