ISSN 1313 - 8820Volume 5, Number 4

December 2013

2013

Scope and policy of the journalAgricultural Science and Technology /AST/ – an International Scientific Journal of Agricultural and Technology Sciences is published in English in one volume of 4 issues per year, as a printed journal and in electronic form. The policy of the journal is to publish original papers, reviews and short communications covering the aspects of agriculture related with life sciences and modern technologies. It will offer opportunities to address the global needs relating to food and environment, health, exploit the technology to provide innovative products and sustainable development. Papers will be considered in aspects of both fundamental and applied science in the areas of Genetics and Breeding, Nutrition and Physiology, Production Systems, Agriculture and Environment and Product Quality and Safety. Other categories closely related to the above topics could be considered by the editors. The detailed information of the journal is available at the website. Proceedings of scientific meetings and conference reports will be considered for special issues.

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Editor-in-Chief

Tsanko YablanskiFaculty of AgricultureTrakia University, Stara ZagoraBulgaria

Co-Editor-in-Chief

Radoslav SlavovFaculty of AgricultureTrakia University, Stara ZagoraBulgaria

Editors and Sections

Genetics and Breeding

Atanas Atanasov (Bulgaria)Ihsan Soysal (Turkey)Max Rothschild (USA)Stoicho Metodiev (Bulgaria)

Nutrition and Physiology

Nikolai Todorov (Bulgaria)Peter Surai (UK)Zervas Georgios (Greece)Ivan Varlyakov (Bulgaria)

Production Systems

Dimitar Pavlov (Bulgaria)Dimitar Panaiotov (Bulgaria)Banko Banev (Bulgaria)Georgy Zhelyazkov (Bulgaria)

Agriculture and Environment

Georgi Petkov (Bulgaria)Ramesh Kanwar (USA)

Product Quality and Safety

Marin Kabakchiev (Bulgaria)Stefan Denev (Bulgaria)Vasil Atanasov (Bulgaria)

English Editor

Yanka Ivanova (Bulgaria)

2013

ISSN 1313 - 8820 Volume 5, Number 4December 2013

Sanitary hygienic assessment of drinking water from underground source at a pig farm

G. Kostadinova*

Department of Applied Ecology and Animal Hygiene, Faculty of Agriculture, Trakia University, 6000 Stara Zagora, Bulgaria*

Abstract. The aim of the study was to investigate and assess the quality of drinking water, obtained from own well source at a pig farm (with an average daily number of reared pigs 532 units), situated in South Bulgaria, Plovdiv Municipality, on the base of main organoleptic, physical, chemical and microbiological indices. For this purpose once a month (from July 2009 to March 2010.) water samples were collected and analyzed by reference methods of Bulgarian State Standard. It was found that the quality of water correspond to the norms concerning organoleptic indices (smell – State 0, norm up to State 2; color – 5°color degrees, norm up to 15°color degrees; visibility – over 65 cm thickness of the water plast through that should clearly seen special text, norm at least 30 cm; taste – no flavor, norm no flavor), chemical indices (pH 7.45÷7.75, norm 6.5–9.5; oxidizability 0.72÷2.16 mg O /L, norm up to 5.0 mg O /L; total hardness 2.82÷4.45 2 2

mg Σqv/L, norm up to 12.0 mg Σqv/L; content of chlorides 20.3÷36.0 mg/l, norm up to 250.0 mg/L, ammonia – not proved, norm up to 0.50 mg/L, nitrites 0.007-0.094 mg/L, norm up to 0.50 mg/L, and nitrates 3.0÷25.0 mg/L, norm up to 50.0 mg/L) and total number of cultivable microorganisms (5 -18 CFU/mL), norm - without significant variability of the index value of the investigated water. The only exception was found for the water temperature, whose maximum values

oexceeded the requirements of the standard (16 C) and number of Escherichia coli (Coliform's test), which values were higher than norm (0 CFU/100 mL), for both indices during five months from the studied period. The deviation in the quality of water in respect content of E. coli suggests anthropogenic pollution on groundwater of the own well with coliform bacteria, containing in manure and urine from the pigs in the farm. Organoleptic, chemical and microbiological characteristics of the sampled water meet the requirements of the Dutch standard for drinking water for pigs. This standard allows higher level of oxygenation of the water and higher levels of ammonium and nitrate content in water compared to Bulgarian standard for humans and animals drinking water. Perhaps time has come, as in the Netherlands to develop a national standard for drinking water, which be applied only to farm animals.

Keywords: pig farm, water, indices, assessment, quality

Abbreviations: BSS – Bulgarian State Standard, CFU – Colony Forming Units, NEc – Number of coliform bacteria (E. coli), TNCM – Total number of cultivable microorganisms

AGRICULTURAL SCIENCE AND TECHNOLOGY, VOL. 5, No 4, pp , 2013448 - 454

Introduction inappropriate management (cleaning, transportation, storage and utilization) can lead to pollution of surface and groundwater in the area of the farm and may lead to serious environmental problems Water resources of the country are unevenly distributed in (MPSC, 1991; Converse, 1993; Ap Dewi, 1994; Cherny, 1997; Lifsey space and time. The average annual volume of water flow is about

3 3 and Jackson, 2005). In EU water pollution with nitrates (in places 19.2 billion m , of which 7.25 billion m are in North Bulgaria and 12.9 3 reached 50 mg/L) in most is associated with animal waste billion m - in Southern Bulgaria. The highest is the water runoff in the

(Voermans, 1993). Webb and Archer (1994) reported that 37% of all high mountain and sparsely populated areas, while in densely cases of water pollution in the UK are caused by wastes from populated plains and hilly areas of Dobrudja and the Upper Trakia livestock farms and 31% of the pollution was significant. valley has a shortage of water resources (Ignatova, 1992). In such

According to data of Iliev et al. (2004), Georgieva et al., (2010, cases, to satisfy drinking and industrial needs of people and farm 2011) and the Annual Report on of Environment in Bulgaria (2012) in animals with water is made to use of groundwater recourses. The many regions of the country ground water contains excessive total amount of stocks of groundwater is estimated at about 6.1

3 3 amounts of nitrates, especially in areas with intensive farming and billion m , of which are used 2.7 billion m . According the conditions plant production. Number of studies (Еnsminger and Parker, 1984; of their formation, these waters are divided into two groups – ground Katsarov et. al., 2003; Geers et al., 2003; Petkov et al., 2004) have and artesian. Greater risk of pollution has as ground watercourses, shown that the intensive technologies applied in pig farms, lead to which are shallower to the surface of the earth. Their regime is generate a large amount of manure wastes that are potential determined by the amount of precipitation and surface water flows. contaminants of the environment. Gomarasca (1996) found that Ground watercourses are connected with surface waters and soils, wastewater from pig farms are the main source of nitrogen pollution and contains both organic matter and microorganisms. Artesian of the water of the rivers Po and Tevere in Italy. Niewolak and Golas waters are formed between two watertight horizons at a greater (2000) established content of NH -N, NO -N, NO -N (above 0.5 4 2 3depth of underground watercourses. These waters are generally mg/L) and a large number of sanitary-indicative microorganisms (E. cleaner than ground watercourses (Jorgensen, 1991; Fewtrell and coli) in the water of the deep water supply wells in the areas of large Bartram, 2001).pig farms.Nowadays, many production units in livestock use water from

Local supply with water from own wells usually used in small their own groundwater sources to satisfy drinking, technological and farms. Sometimes the water from these sources can be hazardous sanitary needs of animals and staff. At the same time, wastes from to human and animal health in the farm, because there are potential livestock production systems (faeces, urine, sewage, litter), by opportunities for its contamination with dangerous substances and

* e-mail: gkostadinova@af.uni-sz.bg

448

449

microorganisms (Nielsen, 1991; Gray, 1996; Cools et al., 2001; 10 m distance in northwest direction from the production buildings Hanselman et al., 2003; Unc and Goss, 2004). Studies in dairy and at 50 m - from the manure storage place. (Kostadinova et al., 2003), pig (Petkov et al., 2004) and poultry Sampling. Water samples for organoleptic, physical and (Stefanova et al., 2012) farms in the country indicate that water chemical analysis were taken monthly from the water source, in quality derived from own drill wells usually meet the quality standard. glass bottle (1 L), and for microbial analysis - in into sterilized glass Nevertheless, these authors noted that risk exists, on the one hand, containers (0.5 L), strictly according to BSS ISO 5667-5 Water when the water contains both nitrate and nitrite, which causes the Quality. The total number of water samples taken was 18. The synergistic effect and on the other hand - by infiltration into the water collected samples were transported to the laboratory ECOLAB at the of pathogenic microorganisms. In this connection interest is the Department of Applied Ecology and Animal Hygiene, Faculty of experience of the Netherlands, where is accepted an standard for Agriculture, Trakia University, Stara Zagora, Bulgaria, in a cool bag

oquality requirements for drinking water only for pigs (DAHSDWSP, (at 4–6 C), and processed for microbiological analysis up to 2 h after 2003). This standard permits higher levels of water contamination of the collection, and for chemical analysis on the next day. some indices in comparison with the Bulgarian standard for drinking Organoleptic (smell, color visibility, taste) and physical water for human beings. This expands the possibilities for use of the (temperature) indices of water were determined in situ, immediately available water resources for livestock. Time will show whether this after sampling water. approach has been correct and adequate. Investigated indices of water and methods of analysis. For the

The aim of the study was to investigate and assess the water purposes of the study were investigated the following indices of the quality from own water source at a pig farm, located in Central South water and was used the following methods for their analysis:Bulgaria, on the base of main organoleptic, physical, chemical and A. Organoleptic indices microbiological indices and its suitability to be used for drinking and · Smell – descriptive in State by intensity of the smell, BSS production purposes in the farm. 17.1.4.01-77;

· Color – visual in color degrees by platinum-cobalt scale, BSS 17.1.4.01-77;

· Visibility – in cm by method of Snellen, BSS 8451-77; Material and methods· Taste – organoleptic (descriptive), BSS 8451-77.

B. Physico-chemical indices Study area. The study was carried out in a pig farm situated at o· Temperature ( C) – with mercury thermometer, BSS 8451-Trilistnik village, Plovdiv Municipality, South Bulgaria, for the period

77; from July 2009 to March 2010. The farm is located at the southern 2 2 · Active reaction (pH), BSS EN ISO 10523;part of the village on area of 2670 m , of which 900 m are built area.

· Permanganate oxidation (mg O /L), BSS EN ISO 8467:01; At the farm there are three production buildings of equal size (length 2

24.00 m, width 9.00 m, height – 3.00 m) situated parallel to each · Total hardness (mg Σqv/L), BSS EN ISO 6059;other in the direction of "East-West". Production buildings are · Chloride content (mg/L), BSS 17.1.4.27; connected by a common hallway with service premises (storage for · Content of ammonium nitrogen (mg/L), BSS ISO 7150-1; feed, fodder kitchen, office, and veterinary office). In the yard of the · Content of nitrite nitrogen (mg/L), BSS ISO 7890-3; farm there is a silo for grain storage, a shed with weighing scales, a · Content of nitrate nitrogen (mg/L), BSS EN 26777. ramp for loading/unloading of animals and service workshop. C. Microbiological indices

During the studied period the average daily number of the pigs · Total number of cultivable microorganisms (TNCM –

reared in the farm was 532 units, into different categories as follows: CFU/mL). It was enumerated by performing serial dilutions (from

sows and litters – 50 units, prenursery pigs – 82 units, nursery pigs – 1:10 to 1:1,000,000) in saline phosphate-buffer which was vortex

123 units, growing/finishing pigs – 277 and boars – 4 units. Sows and agitated for 1 min prior to plating onto Nutrient Agar (BBL, USA)

litters, and boars were reared in individual pens on solid concrete oplates and incubated for 24 h at 30±1 C under normal atmospheric floors, and the other categories of pigs (prenursery, nursery,

conditions. CFU in all plates were enumerated after 24 h. (Smith, growing/finishing pigs) – in group pens (15 – 18 pigs) on slotted

1980; Danon-Moshe et al., 1985).floors, respectively.

· Number of coliform bacteria (E. coli) (NEc – CFU/100 mL). Manure management. Total amount of manure was calculated For the determination of this parameter samples of water were based on the norms for solid and liquid manure, received by a pig diluted as described above but plated onto MacConkey Agar (BBL, from each category (TSDLPCF, 1982; Еnsminger and Parker, 1984). oUSA) plates and incubated at 37±1 C under normal atmospheric Daily manure production in the pig farm was as follow: solid manure

3 3 conditions. CFU in all plates were enumerated after 24 h. (Alef, – 1020 kg (5.8 m ), liquid manure – 1300 L (1.3 m ). Manure from 1991; Jeffery et al., 2004).animals (feces and urine) falls into channels under the slotted floors.

Assessment of water quality. Ecological quality assessment of After filling the channels (up to 5-6 cm below the lower edge of the the tasted water samples was made by comparing the obtained channels), the collected manure discharged into collector channel, results for organoleptic, physico-chemical and microbiological and through it is poured into a settling tank. There the wastes were indices with statutory norms for different indicators in Regulation № separate in solid and liquid fraction. After that the solid fraction was

3 1/2007 to study, use and protection of groundwater and Regulation stored in groundhandling manure storage (400 m capacity) and the № 9/2001 on water quality intended for drinking purposes.3liquid one - in underground concrete tank (150 m capacity). The

The water quality assessment at simultaneous presence in the obtained manure is used to fertilize agricultural land around the farm,

water of nitrates and nitrites was made by the formula (Regulation usually in spring and autumn.

No 9/2001):Water supply. The water in the farm for drinking (about 4000 - -(NO )/LC + (NO )/LC ≤ 1,2 1 3 2L/d) and technological purposes (3000 L/d) was supply from own

- -where (NO ) is content of nitrites in water, mg/L; (NO ) – Content of 2 3well drilling with depth 10 m and flow rate 6 L/s. The well is located at

450

nitrates in water, mg/L; LC – Limit concentration of nitrites in the 1

water, 0.50 mg/L; LC – Limit concentration of nitrates in the water, 2

50 mg/L. This comprehensive index makes it possible to determined synergistic effect of simultaneously content of nitrates and nitrites in the water.

Statystical analysis. The data obtained were analyzed using the Program Statistica 7 for Windows.

Results and discussion

Organoleptic indices. All water samples analyzed for organoleptic indices corresponded to statutory requirements: Smell – water has no smell, so it was assessed with State 0 (at norm up to State 2); Color of the water (expressed in color degrees) was 5° (at norm up to 15° or acceptable for consumers); Visibility of water (thickness of the water layer in cm through that should clearly seen special text) was over 65 cm, at norm at least 30 cm; Taste - the water was no flavor (at norm – no flavor) (BSS 2823-83/1983; Regulation

autumn. According Ignatova (1992) as much as greater is influence № 9/2001). of the air temperature on the temperature of the ground water, as Physic-chemical indices. Results for the studied physic-higher is the risk of their contamination, since the water passed close chemical indices of water are presented in Table 1. to the ground surface. In our previous studies of local water sources Temperature (°C). This parameter of water during study period in dairy farms (Kostadinova et al., 2003), as well as studies of other oscillated between 12.8 and 22.0 °C (Table 1, Figure 1). The water authors on the problem – Petkov et al. (2004) in pig farms and temperature exceed the upper permissible limit for drinking water Stefanova et al. (2012) in poultry farms, showed that the water (16 °C) in July, August, September, October and February. During temperature from own sources with depth between 10 and 15 m the rest months of the controlled period (November, December, varies in much narrower borders (13.5–16.0 °C) than those in this January and March) the water temperature was in norm. The values study, regardless that all these sources are located in close and o f the coefficient of variance revealed the greatest variation of the similar rural regions. In our opinion, the main reason for this temperature in the winter (C = 10.8%), and at least in the summer v difference in the water temperature is the specific characteristics of

(C = 6.4%). This gives reason to assume that the impact of seasonal v each water source (topography, subsoil, depth of the water source).temperature of air on the water temperature of the survey water Active reaction (pH). The active reaction of water varied in very source is greater during in the summer than in the winter and

Table1. Physical and chemical indices of water

* NP – Not Proven, ** According Regulation No 9/2009 and BSS 2823-83 Drinking water (for T °C),*** According DAHSDWSP, (2003), **** Means in the same columns indicated by equal letters are statistically different at Р ≤ 0.05 for aa, bb and at Р ≤ 0.01 for cc

Season

Summer

CV, %

Autumn

CV, %

Winter

CV, %

Avg. for the period

CV, %

Min

Max

Bulgarian Standard**

Netherlands Standard***

b14.0±1.51

10.8

16.5±1.32

8.1

12.8

22.0

6 – 16

-

3.61±0.30

8.3

3.94±0.19

4.8

2.82

4.45

≤ 12.0

5.4 – 9.0

21.1±2.81

13.3

27.7±2.64

9.5

20.3

36.0

≤ 250

≤ 250

c7.48±0.04

0.53

7.62±0.04

0.52

7.45

7.75

6.5 – 9.5

5.0 – 8.0

1.27±0.20

15.7

1.21± 0.15

12.4

0.72

2.16

≤ 5.0

≤ 50

ab20.9±1.34

6.4a14.7±1.08

7.3

c7.70±0.01

0.12

7.68±0.05

0.65

1.16±0.03

2.6

1.22±0.27

22.3

4.02±0.04

1.0

4.20±0.27

6.4

28.9±1.33

4.6

31.0±3.80

12.2

NP*

-

NP

-

0.007±0.005

71.4

0.011±0.008

72.7

19.5±1.00

5.12

15.0±2.87

18.5

Indices

Temperature,Т °С

рН

2009

2010

Oxidizabilityby KMnO ,4

mg O /L2

Totalhardness,mg Σqv/L

Chlorides,mg/L

Ammonia,mg/L

Nitrates,mg/L

Nitrites,mg/L

NP

-

NP

-

NP

NP

≤ 0.50

≤ 1.0

0.062±0.038

61.3

0.026±0.017

66.3

0.007

0.094

≤ 0.50

≤ 0.10

19.7±1.88

9.5

18.1±1.97

10.9

3.0

25.0

≤ 50

≤ 100

Figure 1. Temperature of water, T °C

2522

12.8 12.8

T °C

16 °C

Temperature

norm

norm

6 °C

20

15

10

5

0

Months (2009–2010)

VII VIII IX X XI XII I II III

451

narrow range from 7.45 to 7.75, C =0.52% (Table 1, Figure 2). The when it is negligible. Results found by Kostadinova et al. (2003), v

measured values were in the norm for this index (6.5 – 9.5) stipulated Petkov et al., (2004) and Stefanova et al., (2012) in underground waters in dairy, pig and poultry are in the same range (0.24 – 2.18 mgO /L).2

Total hardness (mg Σqv/L). Water total hardness varied between 2.82 and 4.45 mgΣqv/L, i.e. in the range significantly lower than the permissible level (up to 12.0 mgΣqv/L) (Table 1). These

values c haracterized water as soft water. The standard of the

Netherlands for the quality of drinking water for pigs allow lower levels of variation of this parameter (5.4 – 9.0 mgΣqv/L) (DAHSDWSP, 2003). The values of the index fluctuated in a relatively narrow range, average for the investigated period C =4.8%. Kostadinova et al. (2003), Petkov et al. (2004) and v

Stefanova et al. (2012) reported higher values and larger variation of

the values o f that index (3.8 – 11.6 mgΣqv/L), but within the norms

for drinking water from own sources in dairy, pig and poultry farms, situated in similar neighbor areas in Central Southern Bulgaria. All this shows that the total hardness of groundwater of those regions satisfy quality requirements.

Chloride content (mg/L). Concentrations of chloride in water sampled were low – from 20.3 to 36.0 mg/L with relatively narrow ranges of variations (C vas from 4.6% in summer to 13.3% in v

in Regulation № 9/2001. Similar results (pH between 6.8 and 7.9 winter), (Table 1). All established levels were in the norm – up to 250 units) found and other authors in waters from own underground mg/L, according national (Regulation № 9/2001) and Dutch sources in dairy (Kostadinova et al., 2003), pig (Petkov et al., 2004) legislation (DAHSDWSP, 2003). Obtained results are comparable to and poultry (Stefanova et al., 2012) farms, situated in neighboring those established by Kostadinova et. al. (2003) and Petkov et al. areas. These results indicate that the factors that affect the active (2004) in water from underground sources in dairy and pig farms reaction (pH) of groundwater in the studied regions are the same or (10.5 – 54.5 mg/L), but they are significantly lower than the results very close. Dutch standards for drinking water for pigs allowed lower reported by Stefanova et al. (2012) for underground water source in limits of variation of this indicator (pH=5–8), which may be poultry farms (162.0 – 197.0 mg/L). determined by the specific factors in the country that influence the Content of ammonium nitrogen (mg/L). Neither of the tested values of this index (DAHSDWSP, 2003). samples of water showed no content of ammonium ions (Table 1).

Permanganate oxidation (mg O /L). Values of this index were in 2 Therefore, at permissible level of ammona up to 0.5 mg/L by national the range from 0.72 to 2.16 mg O /L. The established quantities were and up to 1.0 mg/L by the Dutch norms, the analyzed water meets 2

over 2.3 times lower than the permissible limit – up to 5.0 mgO /L the quality requirements. The absence of ammonium ions in the 2

water indicates that no fresh water pollution with organic matter of (Table 1, Figure 3). Consequently, on this indirect indicator animal origin.characterized the water pollution with organic matter, indicates that

Content of nitrite nitrogen (mg/L). During the survey period were the water quality meets the requirements of the standard. The Dutch found low concentrations of nitrite in the water, but with large standard for the quality of drinking water for pigs allow much higher fluctuation between 0.007 and 0.094 mg/L (Table 1, Figure 4). value for this parameter – up to 50.0 mg/L (DAHSDWSP, 2003). The Quantity of nitrite in water varied significantly during the different values of the index range to the greatest extent in the autumn (C = v

seasons of the controlled period, C was from 61.3% in winter to 22.3%), followed by winter (C = 15.7%) and summer (C = 2.6%), v v v

Figure 2. Active reaction (pH) of water

pH

norm

norm

10pH

7.757.45

9.5

6.5

9

8

7

6

5

4

3

2

1

0

Months (2009–2010)

VII VIII IX X XI XII I II III

Figure 3. Oxidizability of the water by KMnO , mg O /L4 2

6 Oxidizability, mg O /L2Oxidizability

norm

5

4

3

2

2.16

0.72

1

0

Months (2009–2010)

VII VIII IX X XI XII I II III

Figure 4. Content of nitrites in water, mg/L

0.6

0.5

0.4

0.3

0.2

0.1

0

0.094

Nitrites

normNitrites, mg/L

-0.1Months (2009–2010)

VII VIII IX X XI XII I II III

452

72.4% in autumn. All values were much lower than the permissible limit (0.5 mg/L) on Bulgarian standard. On this indicator Dutch statdard is stricter than the Bulgarian standard, such as enables quantity of nitrite in water to 0.1 mg/L. The found values do not exceed and this limit. Obviously, the content of nitrites in groundwater used for drinking and production purposes in livestock farms in Central South Bulgaria is not a problem, that affirm and by the results obtained by Kostadinova et al. (2003), Petkov et al. (2004) and Stefanova et al.(2012). These authors reported for levels of nitirites in groundwater of different livestock farms up to 0.13 mg/L.

Content of nitrate nitrogen (mg/L). Nitrate content in water ranged from 3.0 to 25.0 mg/L (Table 1, Figure 5), at permissible level 50.0 mg/L, according Regulation № 9/2001. Dutch norms for

of microorganisms is very small. Significantly greater fluctuation and significantly higher number of cultivable microorganisms in the water from own sources found Kostadinova et al. (2003) in dairy (4-1200 CFU/mL) and Petkov et al. (2004) in pig farms (8-132 CFU/m),

drinking water for pigs admits twice high level of nitrate contents – to 100.0 mg/L than BSS (DAHSDWSP, 2003). In this range are also the results (8.4 – 25.5 mg/L) obtained by Kostadinova et al. (2003) and Petkov et al.(2004) for underground waters in farms for cows and pigs. Significantly higher values (58.0 – 110 mg/L), excessed the permissible concentrations found Stefanova et al. (2012) in groundwater from poultry farms.

Assessment of the water quality at simultaneous presence in the water of nitrates and nitrites. Presence of nitrites and nitrates in groundwater is a criterion for water pollution with nitrogenous organic substances. It is likely the contamination of groundwater be a of manure at the farm when it is stored in a location close to the underground water sources. Assessment on this comprehensive index indicates that the water meets the requirements, since the values of the index range from 0.11 to 0.63, i.e. they are smaller than 1 (C<1), (Figure 6). Therefore, the simultaneous presence of measured concentrations of nitrites and nitrates in the water are not hazardous to farm staff and pigs.

Microbiological indices. Results for the microbiological indices of tested water are presented in Table 2.

Total number of cultivable microorganisms (TNCM – CFU/mL). The TNCM in water varied widely and was between 5 and 18 CFU/mL. The significant variability of numbers of cultivable microorganisms was illustrated by the value of coefficients of variation (C ), average for the investigated period Cv=43.8%. v

Despite the considerable variation of TNCM in the water, which is at variance with the standard (Regulation № 9/2001), it could be assumed that the water meets the requirements as the total number

Figure 5. Content of nitrates in water, mg/L

Months (2009–2010)

VII VIII IX X XI

3

25

XII I II III

60Nitrates, mg/L

Nitrates

norm

50

40

30

20

10

0

Figure 6. Water quality assessment on base of the simultaneous presence of nitrates and nitrites

1.2

1

0.11

0.06

0.63

0.5

0.19

0.8

0.6

0.4

0.2

0

Months (2009–2010)

VII VIII IX X XI XII I II III

( -NO )/LC12-(NO )/LC13

C 1Limit≤

- -*C = (NO )/LC + (NO )/LC 1 (According Regulation No 1/2007)2 1 3 2 ≤

Table 2. Microbiological indices for water qualityassessment from own source at a pig farm

* Not Proven** According Regulation No 9/2009*** According DAHSDWSP, 2003

Months

July

August

September

October

November

December

January

February

March

Avg. forthe period

Cv, %

Min

Max

Bulgarian Standard**

Netherlands Standard***

NP

NP

NP

1.44 ± 1.94

123.7

NP

5

0/100 mL

0/100 mL

5

6

8

8.88 ± 3.89

43.8

5

18

Without significant variability

of the value index of the

investigated water

100 000

18

7

10

9

12

5

1

1

1

NP*

5

5

2009

2010

Total number ofmicroorganisms,

CFU/mL

Number ofEscherichia coli,

CFU/100 mL

453

respectively. Dutch Standard permits very large numbers of develop a standard for drinking water, which be applied only to farm saprophytic microorganisms in drinking water for pigs – up to animals.100000 CFU/mL.

The TNCM in the water varies during different seasons. The number of microorganisms was greatest in the summer (11.7±6.3 Acknowledgments CFU/mL), followed by autumn (8.7±3.3 CFU/mL) and winter (6.3±1.7 CFU/mL). Such dependence established and other This study was implemented within the project “Study and authors who have analyzed water from underground sources in assessment of the ecological status of pig-farm for 50 sows”, ref. No farms for cows (Kostadinova et al.,2003) and pigs (Tihonova and 1E/2009, financed by the Faculty of Agriculture, Trakia University, Alexandrov, 1995; Petkov et al., 2004). With great degree of Stara Zagora, Bulgaria. certainty it may be assumed that the primary factor that influences the number of microorganisms in the water is the level of organic pollution and ambient temperature. Obviously, the higher ambient Referencestemperature during the summer favors the increasing number microorganisms when compared with the winter, when the ambient

Annual Report on State of Environment in Bulgaria, 2012. temperature is lowest.

Executive Environment Agency, Sofia.Number of coliform bacteria (E. coli) (NEc–CFU/100 mL). The

Alef K, 1991. Methodenhandbuch Bodenmikrobiolgie (Aktivitaten-NEc in the water ranged from samples in which no prove any

Biomasse-ifferenzierung). Lendsberg/Lech, Ecomed, Germany, pp. bacteria in the water (October, January, February, March) to 5

13–43.CFU/100 mL (November and December) (Table 2). The results show

Ap Dewi I, 1994. The Use of Animal Waste as a Crop Fertilizer. In: that in 5 of the 9 months of the studied period the water did not meet

Pollution in Livestock Production Systems, CAB INTERNATIONAL, the requirement of quality on this indicator, since the number of the

Wallingford, UK, 309-332.coliform bacteria in 100 ml water was 1 or larger. Presence of

Bulgarian State Standard (BSS) 2823-83, 1983. Drinking water.coliforms in the water is an indicator of fresh fecal pollution. The NEc

Cherny NV, 1997. Hygienic condition of farms and swine health. In: in water showed a very large variability in the different months of the thProceedings of the 9 International Congress in Animal Hygiene, 17-year, which implies sensitivity of that parameter to many and varied

21 August 1997, Helsinki, Finland, 1, 234-237.environmental factors. It is very likely one of the sources of

Converse J, 1993. Global importance of managing livestock waste. groundwater contamination by microorganisms, including with

In: Proceeding of Livestock Waste Management Conference, coliform bacteria to be manure from the pig farm. All this requires that

January 19-20, 1993, St. Paul, Minnesota, USA, 2-7.microbiological indices of water (TNCM and Nec) to be determined

Cools D, Merckx R, Vlassak K and Verhaegen J, 2001. Survival of and controlled.

E. coli and Enterococcus spp. derived from pig slurry in soils of In order to expand opportunities to provide sufficient drinking

different texture. Applied Soil Ecology, 17,1, 53-62.water from own underground water sources for livestock farms,

Danon-Moshe S, Kozareva MN and Paparikova KD, 1985. perhaps time has come, as in the Netherlands to develop a standard Sanitary Microbiology - Methods of Study. Medicine Press, Sofia, pp. for drinking water, which be applied only to farm animals. Before that, 139-165. it is necessary to conduct a thorough scientific research in order to Dutch Animal Health Service Drinking Water Standards for Pigs clarify all aspects of the potential risks to the health and productivity (DAHSDWSP), 2003. Pig International, 33, 10, 14-15.of livestock.Еnsminger ME and Parker RO, 1984. Swine management. In:

thSwine Science, Edited by Еnsminger, M.E. and R.O. Parker, 5 Edition. The interstate Printers & Publishers, Inc., Danville, Illinois,

Conclusion USA, 353-374.Fewtrell L and Bartram J, 2001. Water Quality - Guidelines,

The quality of water from underground water source in the Standards and Health: Assessment of Risk and Risk Management studied pig farm meets the statutory standards on investigated for Water-Related Infectious Diseases, First Edition. World Health organoleptic indices (odor, color, visibility and taste), chemical Organization, Geneva, Switzerland, pp. 44-62.indices (pH, oxidizability, total hardness, content of chlorides, Geers R, Petersen B, Huysmans K, Knura-Deszczka S, De ammonia, nitrites and nitrates) and total number of cultivable Becker M, Gymnich S, Henot D, Hiss S and Sauerwein H, 2003. microorganisms. The only exception was found for the water On farm monitoring of pig welfare by assessment of housing,

management, health records and plasma haptoglobin. Animal temperature, whose maximum values e xceeded the requirements of o Welfare, 12, 4, 643-647.the standard (16 C) and number of Escherichia coli (Coliform's test),

Georgieva N, Yaneva Z and Kostadinova G, 2011. Spatio-which values were higher than norm (0 CFU/100 mL), for both temporal Distribution of Nitrates, Nitrites and Ammonium in indices during five months from the studied period. The deviation in Growndwaters. Ecologica, 18, 64, 623-630.the quality of water in respect content of the E. coli suggests Georgieva N, Yaneva Z and Dospatliev L, 2010. Ecological anthropogenic pollution on groundwater of the well with coliform Monitoring of the Fresh Waters in Stara Zagora Region, Bulgaria. I. bacteria, containing in storage manure at the farm. Quality Analyses of Nitrogen Compounds Contents. Desalination, Organoleptic, chemical and microbiological characteristics of 264, 48-55.the sampled water meet the requirements of the Dutch standard for Gray NF, 1996. Drinking Water quality. Problems and Solutions, drinking water for pigs. This standard allows higher levels of Trinity College, University of Dublin, Ireland, John Wiley & sons, oxygenation of the water and higher levels of ammonium and nitrate Chichester-New York-Brisban-Singapore, 20-48.in water compared to Bulgarian standard for humans and animals Gomarasca MA, 1996. Impact evaluation of agrotechnologies in drinking water. Perhaps time has come, as in the Netherlands to

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Genetics and Breeding

Nutrition and Physiology

Investigation on the possibility to efficiently use Ukrainian cultivars for developing of early winter wheat linesI. Grain productivityN. Tsenov, T. Petrova, E. Tsenova

Combinig ability for grain yield of late maize linesN. Petrovska

Use of recurrent selection in middle late synthetic maize populationI. Results of the first cycle in synthetic “1/2005”N. Petrovska, V. Valkova

Genetic diversity and distance between two Bulgarian local sheep breeds assessed by microsatellite markersS. Georgieva, E. Todorovska, D. Hristova, I. Dimitrova, N. Stancheva, Ts. Yablanski

Testing of new Bulgarian sunflower hybrids under the conditions of North-East BulgariaI. Productivity and traits related to productivityG. Georgiev, P. Peevska, E. Penchev

Comparative morphological study of new Burley tobacco linesT. Radoukova, Y.Dyulgerski

Effect of genotypic and environmental factors on the inheritance of the main characters in chickpea and relationships between themR. Sturzu, T. Nistot, Cr. Melucă, Fl. Bodescu, A. Stoilova

Evaluation of double haploid lines of winter malting barley using selection indicesB. Dyulgerova, D. Valcheva

Evaluation of the combining ability of grain yield of mutant maize lines M. Ilchovska

Comparative study of some biochemical indicators in Karakachan and Copper-Red Shumen sheep breedsG. Angelov, I.Dimitrova, T. Mehmedov, P. Stamberov, N. Stancheva, S. Georgieva, Zh. Nakev

Impaired pancreatic function in mulard ducks with experimental aflatoxicosis I. Valchev, N. Grozeva, D. Kanakov, Ts. Hristov, L. Lazarov, R. Binev, Y. Nikolov

Comparative investigations on feeding efficiency in growing and fattening DanBred and Topigs hybrid pigsG. Ganchev, A. Ilchev

Blood parameters in yearling sheep fed Paulownia (Paulownia spp.) leavesI. Varlyakov, V. Radev, T. Slavov, G. Ganchev

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Changes in some blood parameters in yearling rams fed diets with different protein and lipid levelsV. Radev, T. Slavov, I. Varlyakov

Effect of the sowing norm and nitrogen fertilization on the yield from dry bean (Phaseolus vulgaris L.) cultivar Beslet G. Milev

Evapotranspiration of corn crop for silageR. Bazitov, A. Stoyanova

Productivity and economic traits of winter oilseed rape (Brassica napus var. biennis) under the conditions of DobrudzhaG. Georgiev, G. Georgiev, P. Chamurliyski

Feasibility of the use of heat energy from alternative sources for air conditioning in sows facilityK. Peichev, R. Georgiev

Productivity of green beans, irrigated at different pre-irrigation soil moistureR. Petrova, A. Matev, K. Koumanov, B. Harizanova-Petrova

Comparative assessment of plant resources as substrates for bioshlam productionZ. Shindarska, V. Kirov, G. Kostadinova, B. Baykov

The influence of organic carbon on bioremediation process of wastewater originate from aquaculture with use of microalgae from genera Botryococcus and Scenedesmus I. Sirakov, K. Velichkova, G. Beev, Y. Staykov

Sanitary hygienic assessment of drinking water from underground source at a pig farmG. Kostadinova

Study of bee honey by spectral analysis in the near infrared spectrumI. Zhelyazkova, S. Atanasova , K. Elencheva – Karaneycheva

Comparative GC/MS analysis of lavender (Lavandula angustifolia Mill.) inflorescence and essential oil volatilesT. Zagorcheva, S. Stanev, K. Rusanov, I. Atanassov

Influence of key factors on the time of initial coagulation of cow's milk using milk-clotting enzyme of camel originP. Panayotov, K. Yoanidu, P. Boyanova, B. Milenkov

Production Systems

Agriculture and Environment

Product Quality and Safety

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Мо, Greek = Gr, Georgian = Geor., Japanese = Jа, Chinese = Ch, Arabic = Аr, etc.)The following order in the reference list is recommended:Journal articles: Author(s) surname and initials, year. Title. Full title of the journal, volume, pages. Example:Simm G, Lewis RM, Grundy B and Dingwall WS, 2002. Responses to selection for lean growth in sheep. Animal Science, 74, 39-50Books: Author(s) surname and initials, year. Title. Edition, name of publisher, place of publication. Example: Oldenbroek JK, 1999. Genebanks and the conservation of farm animal genetic resources, Second edition. DLO Institute for Animal Science and Heal th, Netherlands.Book chapter or conference proceedings: Author(s) surname and initials, year. Title. In: Title of the book or of the proceedings followed by the editor(s), volume, pages. Name of publisher, place of publication. Example: Mauff G, Pulverer G, Operkuch W, Hummel K and Hidden C, 1995. C3-variants and diverse phenotypes of unconverted and converted C3. In: Provides of the Biological Fluids (ed. H. Peters), vol. 22, 143-165, Pergamon Press. Oxford, UK.Todorov N and Mitev J, 1995. Effect of level of feeding during dry period, and body condition score on reproductive perfor-

thmance in dairy cows,IX International Conference on Production Diseases in Farm Animals, Sept.11 – 14, Berlin, Germany, p. 302 (Abstr.).Thesis:Penkov D, 2008. Estimation of metabolic energy and true digestibility of amino acids of some feeds in experiments with muscus duck (Carina moshata, L). Thesis for DSc. Agrarian University, Plovdiv, 314 pp.

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Volume 5, Number 4December 2013