




































 

 

 
45 

† Corresponding author 
© 2014 Conscientia Beam. All Rights Reserved. 

SELENO-CYSTINE AFFECTS THE FATTY ACID PROFILE IN IN VITRO 

INCUBATED OVINE RUMINAL FLUID CONTAINING -LINOLENIC ACID 

 

Marian Czauderna1† --- Rozbicka Wieczorek A.J.2 --- Więsyk E.3   
1, 2, 3The Kielanowski Institute of Animal Physiology and Nutrition, Polish Academy of Sciences, Poland 

 

ABSTRACT 

The influence of seleno-cystine (CySe2) added to ovine ruminal fluids containing -linolenic acid (LNA) 

on the profile of fatty acids (FA) was investigated. Fluids were incubated in vitro at 39°C under CO
2 
either 

alone (RF) or with LNA (1.67 mg/ml) or with a combination of LNA with either a low (1.34 μg/ml) 

or high (3.33 μg/ml) level of Se as CySe2. Fluids were removed after 0, 6, 12, 18, 24 hrs of incubation and 

then analyzed to determine FA levels. LNA added to the fluids without/with CySe2 decreased the C180 

concentration for incubation at all times from 6 hrs compared with the RF or the fluid containing CySe2. 

LNA added to the fluids without/with CySe2 decreased the biohydrogenation yield to C18:0. CySe2 added 

to the fluids decreased the C18:0 concentration and the index of the biohydrogenation to C18:0 compared 

with the RF. The higher concentration of CySe2 in the fluids with LNA reduced the accumulation of 

trans11C18:1 for incubation at all times from 18 hrs compared with the fluids with LNA, irrespective of 

the presence of the lower concentration of CySe2. The lowest concentration of trans11C18:1 in the fluids 

with LNA and the higher concentration of CySe2 correlated with the lowest yield of the isomerization of 

LNA into cis9trans11cis15C18:3 and the lowest yield of the initial biohydrogenation of 

cis9trans11cis15C18:3 to trans11cis15C18:2 in the fluids containing LNA and the higher concentration 

of CySe2. CySe2 added to the fluids with LNA decreased the ratio of polyunsaturated FA to saturated FA 

for incubation at all times from 12 hrs compared with the fluids containing LNA. CySe2 in the fluids 

without/with LNA reduced the FA sum in the fluids.  

Keywords: Selenium, -Linolenic acid, Ovine ruminal fluids, Fatty acids, Biohydrogenation, Isomerization.  

 

Contribution/ Originality 

Our original study documents that CySe2 added to the ovine ruminal fluids, irrespective of 

the presence of αLNA, affects concentrations of fatty acids, the capacity of the bacterial 

isomerases and the biohydrogenation yield of unsaturated fatty acids in in vitro incubated ruminal 

fluids compared with the control fluid.  

 

 

Animal Review 
2014 Vol. 1, No. 3, pp. 45-56 
ISSN(e): 2409-6490 
ISSN(p): 2412-3382 
© 2014 Conscientia Beam. All Rights Reserved. 
 

 
 

 



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46 

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1. INTRODUCTION 

A large proportion of selenium (Se) in feedstuffs is present in organic compounds, as seleno-

cysteine (CySe) and especially as seleno-methionine (Se-Met), and forms a part of the amino acid 

structure of protein molecules in the feed [1, 2]. Rumen microorganisms can incorporate CySe 

and Se-Met into their own proteins, but can also reduce an excess of dietary Se-compounds into 

inorganic forms (like selenide or Seo) that are largely unavailable to the ruminant [3, 4]. Ruminal 

microbial Se concentrations were enriched relative to the concentration of Se in rations. Indeed, 

the concentration of Se was significantly higher than that of the ration, whether considered 

relative to diet dry matter (average of 46-fold), nitrogen (average of 11.3-fold) or sulfur 

abundance (average of 26-fold) [5]. Moreover, the enrichment of Se in microbial cells (2-78-fold) 

was greater than the enrichment of either nitrogen (average of 4-fold) or sulfur (average of 1.6-

fold). Ruminal microorganisms reduce much of dietary inorganic Se (as selenite or selenate) to 

unabsorbable elemental Se or inorganic selenide forms. Bacteria are also able to synthesize Se-

Met and CySe, and then these Se-amino acids (Se-AA(s)) are incorporated into microbial proteins. 

Thus, dietary Se finds its way into a form metabolizable by ruminants, and so the way to improve 

the healthiness of ruminant meat and milk by increasing the concentration of CySe and Se-Met 

[6, 7].   

Recent investigations demonstrated that n-3 polyunsaturated fatty acids (PUFA) like -

linolenic acid (LNA) and long-chain PUFA (LPUFA) possessed also several potential health 

benefits, including cancer prevention, decreased atherosclerosis, improved immune response and 

altered fatty acids (FA) and protein metabolism [8-11]. Numerous in vitro and in vivo studies 

documented that ruminant dietary C18-PUFA like linoleic acid (LA) or LNA are direct 

incorporated into the ruminal bacteria, isomerized to other geometrical and positional isomers, 

metabolized into conjugated linoleic acid (CLA) isomers [8, 10, 12-14], as well as 

biohydrogenated to trans11C18:1 (t11C18:1) and finally to C18:0. Indeed, C18-PUFA, especially 

LPUFA, have toxic effects on cellulolytic bacteria and protozoa, act against ruminal lactate 

producers thereby favouring propionate producers [15-17]. Interestingly, unsaturated fatty acids 

(like LNA, LA or cis9C18:1) inhibited bacterial enoyl-acyl carrier protein reductase, an essential 

component of bacterial fatty acid synthesis [18]. Unsaturated fatty acids (UFA) are more toxic 

than saturated fatty acids (SFA) and can inhibit fermentation in the rumen more intensively [19]. 

UFA are especially toxic to G+ bacteria, whereas G- bacteria are less sensitive to fatty acids at the 

same concentration [20]. Biohydrogenation is a microbial pathway in ruminal contents designed 

to reduce unsaturation of lipids found within plant matter, and is likely an evolutionary 

adaptation to protect the microbial population from antimicrobial effects of unsaturated fatty acids 

(UFA) [8, 13].  Current studies indicated that t11C18:1, the intermediate biohydrogenation 

product of αLNA and LA formed in a rumen, may also be of benefit in the prevention of cancer 

due to endogenous conversion of t11C18:1 to cis9,trans11CLA (c9t11CLA) in mammalian tissues 

[21-24]. In our recent studies it was found that the concentration of UFA in a body of animals, as 

well as in membrane of microorganisms cells were positively correlated with the Se level in a diet 

[25-28]. Indeed, Se is an integral component of antioxidant enzymes (e.g. glutathione 



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47 

© 2014 Conscientia Beam. All Rights Reserved. 

peroxidases) that can decrease the risk of PUFA peroxidation in particular [29, 30]. Moreover, 

our recent studies revealed that of selenate or selenite changed concentrations of FAs and CLA 

isomers in particularly in incubated ovine ruminal fluid muscle [23, 24].  

Considering the above, we hypothesized that Se-cystine (CySe2) added to the ovine ruminal 

fluids affected bacterial isomerization of -linolenic acid (LNA) and reduced the 

biohydrogenation of UFA in in vitro incubated ruminal fluid. Therefore, the major objective of the 

current study was to examine the hypothetically effect of CySe2 on the concentration of UFA, 

especially LNA, their geometric and positional isomers, in in vitro incubated ovine ruminal fluids 

containing LNA.  

 

2. MATERIALS AND METHODS 

2.1. Animals and Diets 

Eight ruminally fistulated adult sheep received a mixed diet comprising grass hay, barley, 

molasses, soybean meal and minerals and vitamins, at 500, 299.5, 100, 91 and 9.5 g/kg dry matter 

respectively, fed in equal meals of 500 g at 8.00 and 16.00 h [23]. Ruminal digesta samples were 

taken before feeding in the morning from each sheep. The ruminal fluid was kept at 39C and 

strained through linen cloth before use.   

 

2.2. Reagents 

CySe2, LNA and fatty acid methyl ester standards were purchased from Sigma (Poole, 

Dorset, UK). Other reagents were of analytical grade and were from POCh (Gliwice, Poland). 

Water used for the preparation of mobile phases and chemical reagents was prepared using an 

ElixTM water purification system (Millipore).  

 

2.3. Incubations with Ruminal Fluid in Vitro 

Strained ruminal fluids (SRF) were incubated either alone or with a combination of LNA 

and two concentrations of CySe2
 to determine the interactions in the metabolism of LNA. All in 

vitro experiments were performed on four different days using samples withdrawn from eight 

different sheep. 

In general, 1 ml of strained ruminal fluid was added under CO2 to Pyrex tubes (120 x 11 mm) 

containing 0.2 ml of water solution containing either alone (the positive control groups) or with a 

combination of LNA, a low (l) or high (h) level of CySe2 (Table 1). The tubes with the ovine 

ruminal fluids were incubated at 39°C. The tubes were removed after 0, 6, 12, 18 or 24 hrs of in 

vitro incubation, heated to inactivate ruminal microbial enzymes for 9–10 min in a block heater at 

100°C and stored at –20°C before being submitted for the quantification of FA. The FA were 

extracted, methylated and analyzed as described below.  

 

2.4. Fatty Acid Extraction and Preparation of Fatty Acid Methyl Esters (FAME) 

The method of alkaline saponification and extraction was as described previously [24]. 

Derivatization of the extracted free fatty acids to FAME was carried out using a procedure that 



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48 

© 2014 Conscientia Beam. All Rights Reserved. 

contained mild, base- and acid-catalyzed methylation steps that minimized positional and 

geometrical isomerization of UFA [24]. FAME were then analysed using gas chromatography 

(GC) according to Czauderna, et al. [24]. The analyses of all FAME in ruminal fluid samples 

were performed on a SHIMADZU GC-MS QP2010 Plus EI equipped with a BPX70 fused silica 

capillary column (120 m x 0.25 mm i.d. x 0.25 µm film thickness; SHIM-POL), a quadrupole mass 

selective detector (Model 5973N) and an injection port. FAME identification was validated based 

on the electron impact ionization spectra of FAME and compared with authentic FAME 

standards and the NIST 2007 reference mass spectra library. 

 

2.5. Statistical Analyses 

Statistical analyses were performed using the Statistica software package (StatSoft, Version 

10, 2010). Statistical analyses of the effects of LNA and CySe2 on the concentrations of selected 

FA in in vitro incubated ruminal fluids were conducted using the non-parametric Mann-Whitney 

U test. The results are presented as the means of the individually analyzed ruminal fluid samples. 

Mean values in the columns with different superscripts are significantly different at a,bP < 0.05 

and A,BP < 0.01. 

 

3. RESULTS 

3.1. The Influence of CySe2 on the Accumulation of Selected Saturated Fatty Acids (SFA) 

in in Vitro Incubated Ruminal Fluids with  LNA      

Although factors altering the microbial population and ruminal fermentation are undoubtedly 

keys to controlling the yield of the biohydrogenation and synthesis of positional or geometric 

isomers of fatty acids very few studies have directly associated production of fatty acid isomers 

and their products of the biohydrogenation in ruminal fluids enriched in Se-compounds [12-14, 

23, 24, 31, 32]. Therefore, in vitro studies were conducted to determine the effect of CySe2 on the 

concentrations of selected fatty acids in ovine ruminal fluids containing extra LNA. The results 

summarised in Table 2 documented that the addition of LNA to the incubated ruminal fluids, 

irrespective of the presence of CySe2, decreased the concentration of C180 from 6 to 24 hrs of in 

vitro incubation compared with the control ruminal fluid (RF) or the fluid containing only CySe2, 

regardless of its concentration. On the other hand, CySe2 added to the ruminal fluids revealed 

minute influence on the accumulation of C18:0 compared with the RF; indeed, CySe2, in dose 

dependent manner, slightly reduced the concentration of C18:0 from 12 to 24 hrs compared with 

the RF. Of interest is the observation that the addition of LNA to the fluids with CySe2, 

irrespective of its concentration, slightly stimulated the accumulation of C14:0 from 6 to 24 hrs 

compared with the RF and other ruminal fluids with LNA or CySe2. Moreover, LNA added to 

the ruminal fluid, irrespective of the presence of CySe2, stimulated the accumulation of C20:0 and 

C22:0 especially from 18 to 24 hrs of incubation compared with the RF and the fluids with CySe2. 

On the other hand, all additives in the incubated fluids revealed negligible influence on the 

concentrations of C15:0 and C16:0 as well as the concentration sum of all assayed SFA (SFA) 

compared with the RF.  



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© 2014 Conscientia Beam. All Rights Reserved. 

3.2. The Influence of CySe2 on the Accumulation of Selected Mono- and Poly-Unsaturated 

Fatty Acids in in Vitro Incubated Ruminal Fluids with LNA      

As can be seen from the results summarized in Table 3, all additives in the incubated ruminal 

fluids revealed negligible influence on the concentrations of c9C14:1 and c9C16:1 compared with 

the RF. Similarly, the addition of CySe2 to the fluids enriched in LNA revealed negligible 

influence on the concentrations of c9C18:1 and t11C18:1 (TVA) as well as the concentration sums 

of cMUFA and tC18:1 compared with the fluids containing only LNA. The concentrations of 

c9C18:1, t11C18:1 and cMUFA were higher from 6 to 24 hrs of incubation in the fluids with 

CySe2 and LNA than in the fluids containing only CySe2.   

As can be seen from results summarized in Table 4, CySe2 or/and LNA added to the 

ruminal fluids affected the concentrations of PUFA in in vitro incubated ruminal fluids. The 

concentration of t11c15C18:2 (tcC18:2) increased throughout the incubation in the ruminal fluids 

enriched in LNA, whereas decreased in the RF and the fluids containing only CySe2. The 

addition of CySe2 to the fluids with LNA reduced the concentrations of tcC18:2 and 

c9t11c15C18:3 (ctcC18:3) from 18 to 24 hrs of incubation compared with the fluids containing only 

LNA. The concentrations of ctcC18:3, t9t11c15C18:3 (ttcC18:3) and LNA were quantitatively 

detected in the fluids containing LNA; the addition of CySe2 to the fluids with LNA revealed 

negligible influence on the concentrations of ttcC18:3 and LNA as well as on the concentration 

sum of PUFA (PUFA) compared with the fluids with LNA.  The addition of CySe2 to the 

ruminal fluids with LNA slightly decreased the concentration ratio of PUFA to SFA 

(PUFA/SFA) from 12 to 24 hrs of incubation compared with the fluids with only LNA. 

Moreover, CySe2 added to the ruminal fluids without or with LNA, decreased the concentration 

sum of all assayed fatty acids (FA) in incubated fluids compared with the RF and the fluids with 

only LNA, respectively. As observed from the obtained results (Table 4), LNA or/and CySe2 

added to the ruminal fluids changes the indexes of the initial, intermediate and final 

biohydrogenation in incubated fluids compared with the RF.    

 

4. DISCUSSION 

Wina, et al. [33] documented that the contents of the major ruminal microorganisms, 

including protozoa, were found to be kept at higher levels than after 24 hrs in in vitro incubation. 

Considering the above, the in vitro incubation periods in the current investigation were set to 6, 

12, 18 and 24 hrs to determine the influence of CySe2 and/or LNA on the concentration of FA 

in incubated ovine ruminal fluids. Interestingly, the concentration of protozoa in incubated fluids 

was highest at 12 hrs of in vitro incubation, whereas only slightly lower (15%) after 24 hrs of 

incubation compared with the initial concentration of protozoa [33]. Our studies and other 

investigations have shown that pH of ruminal fluids containing LNA slightly decreases after 24 

hrs of in vitro incubation of ovine fluids; the pH value is reduced by approx. 0.5 [34, 35].    

Ruminal microorganisms may use the biohydrogenation as a method of defence against 

toxicity of UFA, especially PUFA (like LNA or LA). Therefore, the first step of the 

biohydrogenation consists of the enzymatic bacterial isomerisation, which turns cis bonds into 



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© 2014 Conscientia Beam. All Rights Reserved. 

trans bonds. This isomerisation step is rapid compared with the biohydrogenation step [8]. As 

can be seen from data summarized in Table 2, LNA added to the ruminal fluids, irrespective of 

the presence of CySe2, decreased the yield of the final biohydrogenation (fBH) to C18:0. 

Considering the above, we argue that LNA decreased the activity of group B bacteria in a 

rumen [36]. Indeed, group A included the ruminal bacteria that hydrogenate PUFA (e.g.  LNA 

or LA) into t11C18:1 where their effect was thought to end; on the other hand, group B ruminal 

bacteria were thought to be capable of converting the same PUFA as those of group A bacteria, 

but they are more capable of biohydrogenating a wider range of fatty acids, ending with C18:0 

flow from the rumen [36]. 

As consequence, the addition of LNA to the fluids, regardless of the presence of CySe2, 

revealed negligible influence on the concentration of C14:0, C15:0 and C16:0 in incubated fluids. 

Indeed, these saturated fatty acids are not the products of the fBH [8]. The current results 

documented that added CySe2, in dose depended manner, reduced the formation of C22:0 in the 

fluids containing LNA compared with the fluids with only LNA from 18 to 24 hrs of the 

incubation; so, our results suggested that added CySe2 decreased the capacity of the elongation of 

saturated fatty acids in the incubated ruminal fluids enriched in LNA. Our investigations 

revealed that CySe2 added to the fluids without or with LNA reduced the synthesis of fatty acids 

or/and increased their oxidation. Therefore, the concentration of FA was lower in the incubated 

fluids containing CySe2 without or with LNA than in the RF or the ruminal fluids with LNA.                

Our current studies documented that the higher concentration of CySe2 in the ruminal fluids 

enriched in LNA reduced the accumulation of t11C18:1 from 18 to 24 hrs of incubation 

compared with the fluids containing LNA, irrespective of the presence of the lower 

concentration of CySe2 (Table 3); t11C18:0 is the product of isomerization and/or the 

intermediate biohydrogenation of unsaturated fatty acids (C18:2), like t11c15C18:2 [25]. Indeed, 

the higher concentration of CySe2 in the ruminal fluids containing LNA most efficiently reduced 

the index of the initial biohydrogenation of c9t11c15C18:3 and t9t11c15C18:3 to t11c15C18:2 from 

18 to 24 hrs of incubation (Table 4). Moreover, the lowest concentration of t11C18:1 in the fluids 

with LNA and the higher concentration of CySe2 well correlated with the lowest yield of the 

bacterial isomerization of LNA into c9t11c15C18:3 (ctcC18:3) and the lowest yield of the initial 

biohydrogenation of ctcC18:3 to t11c15C18:2 (tcC18:2) in the incubated fluids containing LNA 

and the higher concentration of CySe2 (Table 4). Therefore, the concentration of ctcC18:3 and 

tcC18:2 in the fluids enriched in LNA and the higher concentration of CySe2 was lower than in 

the fluids with LNA, irrespective of the presence of the lower concentration of CySe2, from 12 to 

24 hrs of the incubation. Considering the above results, we argued that CySe2 added to the 

incubated fluids with LNA reduced the yield of the bacterial isomerization of LNA (Table 3) as 

well as the initial biohydrogenation of ctcC18:3 in dose dependent manner from 18 to 24 hrs of the 

incubation (Table 4). Moreover, we suggest that LNA added to the incubated fluids, irrespective 

of the presence of CySe2, reduced the capacity of the final biohydrogenation to C18:0 compared 

with the RF (Tables 2 and 4). Moreover, CySe2 added to the ruminal fluids decreased the 

concentration of C18:0 as well as the index of the final biohydrogenation to C18:0 compared with 



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the RF. Current observations are also consistent with our recent in vitro studies that have 

reported that selenate or selenite added to the ruminal fluids containing LNA reduced the 

capacity of the bacterial isomerization of C18-PUFA and lowered the yield of the final 

biohydrogenation to C18:0 compared with the fluids with only LNA [24]. Interestingly, LNA 

or/and CySe2 added to the ruminal fluids revealed an inconsistent effect on the index values of the 

intermediate biohydrogenation (Table 4)  involved in the formation of t11C18:1 (TVA). We 

suggest that this inconsistent effect may be due to the fact that t11C18:1 is the substrate which is 

consumed during the final biohydrogenation. Generally, the index values of the intermediate 

biohydrogenation to t11C18:1 were higher in the fluids containing LNA and the higher or lower 

concentration of CySe2 compared with the ruminal fluids with only LNA. On the other hand, our 

results suggest that the higher concentration of CySe2 in the fluids with LNA usually stimulated 

the yield of elongation of c9c12C18:2 to c13c16C22:2 (i.e. the unsaturated fatty acids) compared 

with the incubated fluids containing only LNA. Surprisingly, CySe2 added to the ruminal fluids 

with LNA decreased the concentration ratio of PUFA to SFA (PUFA/SFA) for incubation 

at all times from 12 hrs compared with the fluid containing LNA (Table 4).      

 

5. CONCLUSION  

In conclusion, CySe2, especially the higher concentration of CySe2, reduced the capacity of the 

biohydrogenation of UFA to C18:0 in the ruminal fluids. Moreover, the higher concentration of 

CySe2 in the fluids with LNA most effectively reduced the yield of the bacterial isomerization of 

LNA to c9t11c15C18:3 and the formation of t11C18:1. Our investigations revealed that CySe2 

added to the fluids without or with LNA reduced the accumulation of fatty acids in in vitro 

incubated ruminal fluids.  

 

6. ACKNOWLEDGEMENTS 

Excellent sheep care and courteous assistance throughout the study were provided by Mrs B. 

Domańska and Mrs G. Oktaba.   

 

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Animal Review, 2014, 1(3): 45-56 

 

 
54 

© 2014 Conscientia Beam. All Rights Reserved. 

Table-1. The scheme of in vitro experiments on ovine ruminal fluids1 

 
Group 
 

 
   Additive 

     Additive 
  concentration 

Number (n) of fluid 
samples individually 
analysed in every time2    

   RF3      Water              -               n=6 

   L       LNA     1.67 mg/ml               n=6 

   Sel      CySe2     1.34 g/ml               n=5 

   Seh      CySe2     3.33 g/ml               n=5 

   L-Sel 
     CySe2 

     LNA         
    1.33 g/ml 
    1.67 mg/ml 

              n=6 
               

  L-Seh 
 

     CySe2 

     LNA 
    3.33 g/ml 
    1.67 mg/ml 

              n=6 

1 1 ml of ovine ruminal fluids was added to 0.2 ml of water solution containing either solely -linolenic acid (LNA), seleno-cystine (CySe2) 
at different concentrations (l - low; h - high), or their mutual combinations  
2 n – ovine ruminal fluid samples collected from n sheep at different times and individually analyzed. All results in Tables 2-4 are mean 
values from n fluid samples individually analysed in every time    
3 RF = the negative control group (in vitro incubated 1ml of the ovine ruminal fluid with 0.2 ml of water (the control ruminal fluid) 

 

Table-2. Effects1 of two levels (low, l; high, h) of Se-cystine (CySe2) on metabolism of LNA and the concentration of 

selected saturated fatty acids (SFA) and the concentration sum of SFA (SFA) (g/ml) in in vitro incubated ruminal fluids     

Group 
   and 
  in vitro 
  incubation 
  time,  hrs    

C
1
4

:0
 

  
C

1
5

:0
 

  
  

  
 

  
  

  
C

1
6

:0
  

   
 

   
  

  

  
C

1
8

:0
 

  
C

2
0

:0
 

 C
2
2

:0
 

 
S

F
A

 

RF2        0      3.8 

L            0     3.4a 

Sel         0      3.5a  
Seh        0      3.7a    
L-Sel     0      3.7a 

L-Seh    0      3.7a 

   23.5 

   24.0a 

   24.7a 

   24.2a 

   24.6a 

   24.7a 

66       
    72a 

66a 

       69a   
74a 

73a        

    93 

    99a 

    97a 

    94a 

   100a 

   100a 

 -3 

 - 
 - 
 - 
 - 
 - 

4.3 

0.7Bb 

4.5a 

4.6a 

0.8b 

  - 

191 

199ab 

195ab 

195ab 

203ab 

202ab 

RF        6       4.4 

L           6       5.1ab 

Sel        6       4.5a 

Seh       6       4.6a 

L-Sel    6       5.4b  
L-Seh    6      5.3ab  

   25.9 

   25.6a 

   24.4a 

   24.6a 

   25.8a 

   25.2a 

79 

99ab 

76a 

     77a    
   103b 

  98ab        

   105 

   101a 

   106a 

   105a 

   104a 

   100a 

0.9 

7.8B 

 - 
 - 
 - 
 - 

4.9 

1.4b 

4.7a 

4.7a 

  - 
2.7b 

220 

239abc 

216abc 

216abc 

238bc 

231abc 

RF       12      5.4 

L         12      5.5b 

Sel       12      5.3ab 

Seh      12      5.3ab 

L-Sel   12      5.9b  
L-Seh  12      6.4b 

   25.6 

   25.3a 

   24.7a 

   25.0a 

   26.6a 

   25.3a 

  94      
 102ab 

90ab 

91ab     

 106b     
  104b       

   127 

   101a 

   121b 

   119ab 

   104ab 

   102a 

1.1 

6.3B 

1.0Aa 

  - 
10.9B 

 4.8Bb 

3.2 

2.8b 

4.2a 

4.4a 

4.4a 

2.6b 

256 

243bc 

246bc 

244bc 

258bcd 

245bcd 

RF       18      5.7 

L         18       5.9b 

Sel       18      6.1b 

Seh      18      5.9b 

L-Sel   18      6.3b 

L-Seh  18      6.4b 

   25.1 

   25.8a 

   25.6a 

   24.7a 

   25.2a 

   25.6a 

   95     
 105b 

   98ab 

    96ab    
 107b 

 111b       

   132 

   100a 

   130b 

   124ab 

   104ab 

   102a 

1.0 

4.4B 

1.0Aa 

1.0Aa 

8.4B 

8.9B 

4.5 

9.0c 

4.4a 

4.5a 

6.3ac 

6.1ac 

263 

250cd 

265cd 

257cd 

257cd 

260cd 

RF       24      5.8 

L            24      6.4b 

Sel       24      6.2b 

Seh      24      6.0b   
L-Sel   24      6.5b 

L-Seh  24      7.0b 

   26.9 

   26.3a 

   25.4a 

   24.6a 

   24.4a 

   26.5a 

 102          
 106b 

 100ab 

99ab        
106b       
111b        

   142 

   101a 

   135b 

   131ab 

   101a 

   106ab 

2.0 

6.8B 

1.9Aa 

1.8A 

3.7ABb 

3.9ABb 

3.8 

 12.8Ac 

   4.1b 

   3.4ab 

 10.3c 

   5.9a 

283 

260cd 

273cd 

266cd 

252bcd 

260cd 

1 means in columns with the different letters are significantly different at (a,b)P < 0.05 or at  (A,B)P < 0.01                                     
2 RF – in vitro incubated ruminal fluids without the additives (the control ruminal fluid) 
3 below the quantification limit (LOQ); LOQ was defined as 10 times the average noise level (LOQ was determined according to Lin and 
McKeon [37]) 



Animal Review, 2014, 1(3): 45-56 

 

 
55 

© 2014 Conscientia Beam. All Rights Reserved. 

Table-3. Effects1 of two levels (low, l; high, h) of Se-cystine (CySe2) on metabolism of LNA and the concentration of 

selected monounsaturated fatty acids (MUFA) and the concentration sums of cMUFA (cMUFA)2 and tC18:1 (tC18:1)3 

(g/ml) in in vitro incubated ruminal fluids   

Group 
   and 
  in vitro 
  incubation 
  time,  hrs  c9

C
1
4
:1

 

 c
9
C

1
6
:1

 

  
  
  
 

  
 

c9
C

1
8
:1

  
   
 

   

  
  

 t
9
C

1
8
:1

 

t1
1
C

2
1
8
:

1
 


cM

U
F

A
 


tC

1
8
:1

 

RF          0     4.5 

L            0     4.4a 

Sel         0     4.4a  
Seh        0     4.6ab    
L-Sel     0     4.5a 

L-Seh    0     4.5ab 

   4.5 

   1.4b 

   4.4ad 

   4.6ad 

   2.5ab 

   2.6ab 

  7.7 

   -4 

   7.6a 

  8.2a 

  2.8Ab 

  2.5Ab 

  3.8 

  2.9a 

  6.0b 

  3.9ab 

    - 

    - 

 1.9 

   - 
   - 
 2.2a 

   - 
   - 

16.7 

  5.8Bb 

16.5a 

17.4a 

  9.8b 

  9.6b 

 5.8 

 2.9Ab 

 6.0a 

 6.1a 

   - 
   - 

RF         6     5.5 

L           6      6.9ab 

Sel         6      5.3ab 

Seh        6      5.5ab 

L-Sel     6      7.1b  
L-Seh    6      6.9ab  

   5.5 

   6.1cd 

   5.2cd 

   5.2cd 

   6.4cd 

   6.0cd 

  8.4 

16.1Bc 

  7.3a 

  7.8a 

13.9Bc 

14.7Bc 

  5.3 

    - 

   7.6b 

   8.2bc 

    - 
   3.1a 

 2.3 

 7.3b 

   - 
   - 
 9.3Ab 

 3.2a  

19.4 

29.1A 

17.8a 

18.5a 

27.4Ac 

27.6Ac 

 7.6 

 7.3a 

 7.6a 

 8.2a 

 9.3a 

 6.3a 

RF       12      6.0 

L         12      7.2b 

Sel       12      5.8ab 

Seh      12      6.0ab 

L-Sel   12     7.5b  
L-Seh  12      7.2b 

   6.4 

   4.8ad 

   6.0cd 

   6.1cd 

   6.8cd 

   6.9cd 

  6.3 

16.8Bc 

  5.9a 

  6.8a 

17.3Bc 

16.9Bc 

   6.6 

    - 
   6.2b 

   7.2b 

    - 
    - 

  4.0 

  9.2b 

  4.2a 

  4.0a 

   9.9b 

  9.4a 

18.7 

28.8Cc 

17.7a 

18.9a 

31.7Cc 

31.0Cc 

10.5 

  9.2a 

10.4a 

11.2a 

  9.9a 

  9.4a 

RF       18      6.2 

L         18      7.5b 

Sel       18      6.5ab 

Seh      18      6.3ab 

L-Sel   18      7.6b 

L-Seh  18      7.5b 

   7.7 

   7.7cd 

   6.6cd 

   6.3cd 

   7.0cd 

   7.1cd 

  2.5 

17.2Bc 

  3.1Ab 

  3.9Ab 

17.3Bc 

14.4Bc 

   5.9 

   2.1a 

   6.2b 

   6.8b 

    - 
   2.6a 

  3.9 

15.1Bc 

  4.3a 

  5.3a 

15.5Bc 

14.1Bc 

16.5 

32.4Cc 

16.2a 

16.5a 

31.9Cc 

29.0Cc 

 9.8 

  17.2Ba 

110.4a 

  12.0a 

  15.5Ba 

  16.7Ba 

RF       24      6.7 

L            24      8.0b 

Sel       24     6.4ab 

Seh      24      6.4ab   
L-Sel   24     7.8b 

L-Seh   24     8.0b 

   9.2 

   7.0cd 

   6.6cd 

   6.4cd 

   7.5cd 

   7.3cd 

  1.0 

17.0Bc 

  1.9Ab 

  2.6Ab 

16.3Bc 

11.9Bc 

   6.2 

   4.6ab 

   3.7a 

   3.1a 

   3.0a 

 15.1d 

  1.1 

19.8Bc 

  5.7ab 

  6.2b 

19.8Bc 

14.9Bc 

16.9 

   32.0Cc 

   14.9a 

   15.4a 

   31.6Cc 

   27.1Ac 

 7.3 

  24.4Bd 

    9.3a 

    9.2a 

  22.8Bd 

  30.0Ccd 

1 means in columns with the different letters are significantly different at (a,b)P < 0.05 or at (A,B)P < 0.01; all abbreviations as in Table 2                        

2 cMUFA = c9C14:1 + c9C16:1 + c9C18:1  
3 tC18:1 =  t9C18:1 + t11C18:1 (TVA) 
4 below the quantification limit (LOQ) 

 

 

 

 

 

 

 

 

 

 

 

 

 



Animal Review, 2014, 1(3): 45-56 

 

 
56 

© 2014 Conscientia Beam. All Rights Reserved. 

Table-4. Effects1 of two levels (low, l; high, h) of Se-cystine (CySe2) on metabolism of LNA and the concentrations of 

t11c15C18:2 (tcC18:2), c9t11c15C18:3 (ctcC18:3), t9t11c15C18:3 (ttcC18:3), c13c16C22:2 (C22:2), the concentration sums of 

PUFA (PUFA) and all assayed fatty acids (FA), the concentration ratio of PUFA to SFA (PUFA/SFA) and the 

indexes of the initial, intermediate and final biohydrogenation in in vitro incubated ruminal fluids  

 

 

1 means in columns with the different letter are significantly different at (a,b)P < 0.05 or at (A,B)P < 0.01; all abbreviations as in Table 2   
2 below the quantification limit (LOQ) 
3 the index of the initial biohydrogenation of c9t11c15C18:3 (ctcC18:3) and t9t11c15C18:3 (ttcC18:3) to t11c15C18:2 (tcC18:2) 

(indextcC18:2 =     tcC18:2/(tcC18:2 + ctcC18:3 + ttcC18:3) 
4 the index of the intermediate biohydrogenation of t11c15C18:2 to t11C18:1 (TVA) (indexTVA= t11C18:1/(t11C18:1+t11c15C18:2) 

5 the index of the final biohydrogenation of t11C18:1 to C18:0 (indexC18:0 = C18:0/(C18:0+t11C18:1) 

 

 

 

 

 

 

 

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