INVESTIGATION ON EFFECTS OF HYPERTONIC SODIUM BICARBONATE WITH AND WITHOUT LACTATED RINGER’s SOLUTION ON POTASSIUM COMPENSATION, REHYDRATION STATUS, ENDOCRINAL RESPONSE AND ACIDE-BASE BALANCE IN CALVES WITH METABOLIC ACIDOSIS

Bulent ELITOK
Afyon Kocatepe University, Faculty of Veterinary Medicine, Department of Internal Medicine, 03200-Afyonkarahisar/
Turkey, E-mail: elitok1969@hotmail.com

Abstract. The study was carried out on a total of 40 calves. Twenty calves (n=20) which were treated
with hypertonic sodium bicarbonate in physiological saline served as the Study Group. The remaining
20 calves, which were treated with sodium bicarbonate in physiological saline plus lactated Ringer’s
solution, served as the Control Group. During the study, animals received fresh milk up to 10% of their
body weight, but no antibiotics were used. All of the animals were tested for clinical findings such as
temperature, pulsation and respiration rates, dehydration findings, hematological findings such as white
blood cell, red blood cell, hemoglobin, packed cell volume, mean corpuscular volume, mean corpuscular
hemoglobin, biochemical findings such as aspartate aminotransferase, gamma glutamyl transferase,
total protein, albumine, urea, creatinine, glucose, lactate and blood gas findings, such as total carbon
dioxide, partial carbon dioxide and electrolytes such as potassium, sodium bicarbonate, chlorine and
calcium along with base excess. All the measurements were taken before the study began and at the 1st,
3rd and 7th hours after the start of the study. The results of the study showed that metabolic acidosis in
calves with diarrhea was treated more quickly with lactated Ringer’s solution than with physiological
saline, although the hyperosmolality and hypernatremic states were corrected with both solutions. Consequently,
lactated Ringer’s solution supplied a better clinical response than that of physiologic serum,
and it is considered that practitioners should add it to fluid therapy for more rapid physiological correction
of calves with metabolic acidosis.
Key words: Calves; Newborn animals; Metabolic acidosis; Fluid therapy; Electrolyte balance.

INTRODUCTION

Neonatal calf diarrhea remains the most common cause of morbidity and mortality at pre-weaned
dairy calves worldwide. This complex disease can be triggered by both infectious and non-infectious
causes. The four most important enteropathogens leading to neonatal dairy calf diarrhea are Escherichia
coli, rota and coronavirus, and Cryptosporidium parvum (Maganck, V. et al. 2014)
Fluid, electrolyte, and nutrient deficiencies are crucial to preserve diarrhea in the calves (Hall et al.
1992). However, dehydration due to diarrhea is the result of the fluid loss from the extracellular compartment.
This fluid loss is compensated by the passage of the intracellular fluid to the extracellular fluid,
i.e., the plasma. If Na+ ions lost by feces in diarrhea cannot be compensated, the amount of body fluids
decreases. Thus dehydration and hypovolemic shock develop in advanced conditions (Roussel, A.J.
1993; Lorenz, I. and Kee, J. et al. 2007). A significant amount of total Na+, K+, Cl- and HCO3
- is lost with
diarrhea and the pH of this electrolyte balance disorder resulting in diarrhea is low (Radostits, O.M. et al.
2006; Kee, J. et al. 2010) . In the case of diarrhea, plasma K+ ions increase in the extracellular fluid, but
the decrease in the cell disrupts the potential of the cell membranes and this is the main cause of death,
particularly by affecting myocardium (Foster, D., Smith, G. 2009; Cho, Y. et al. 2013). If losses continue
to increase and become uncompensated, systemic effects of dehydration and metabolic acidosis
are formed (Hunt, E. 1992; Radostits, O.M. et al. et al. 2006). The measurement of blood gases is very
important in terms of understanding the condition and severity of the disease. Blood gas parameters;
PaO2, oxygenation; PaCO2, alveolar ventilation; PaO2 and PaCO2 together, gas exchange; pH, PaCO2,
and HCO3- are very valuable parameters in determining acidosis and alkalosis status (Radostits, O.M.
et al. 2006; Sarnaik, A. and Heidmann, S. 2007; Emiralioğlu, M., Özçelik, U. 2014).
Our aim at this study was to found out the effects of serum physiological, lactated Ringer’s solution plus
hypertonic 8.4% NaHCO3
- on the blood parameters, with regard to the treatment of liquid-electrolyte and acidbase
balance disorders of neonatal diarrheal calves, and provide new scientific information to clinicians and
veterinary medicine.

MATERIALS AND METHODS

Materials
The study was carried out on a total of 40 calves with diarrhea. Twenty calves were treated with
Sodium Bicarbonate (HCO3
-) in physiological saline (PS) and served as the Study Group (SG) and the
remaining 20 calves were treated with HCO3
- in PS plus along with Lactated Ringer Solution (LRs) and
formed the Control Group (CG). All the measurements were taken before the study began and at the
first, third and seventh hours after the start of the study. During the study, all the animals were given
fresh milk up to 10% of their body weight, and no antibiotics were used.
The present study was carried out in accordance with the ethical rules of the Ethics Committee of
Afyon Kocatepe University, with the reference number AKUHADYEK 197-17, and was supported by
16.KARIYER.120 reference number and the Afyon Kocatepe University Scientific Research Projects
Coordination Unit (BAPK).
Method
In this study, clinical findings such as body temperature, respiration and heart rates, dehydration,
appetite control and live weight gain were recorded. Hematological parameters such as total leukocyte
(WBC), erythrocyte (RBC), mean corpuscular volume (MCV), hematocrit (HCT), mean cell hemoglobin
(MCH), mean corpuscular hemoglobin concentration (MCHC) were measured with a hemocell counter.
In the study, measurements of blood serum parameters such as aspartate aminotranferase (AST), gamma
glutamyl transferase (GGT), albumin (ALB), total protein (TP), glucose (GLU), urea (UREA) and creatinine
(CREA) were made in the Roche Cobas C111 model autoanalyzer using commericial kits.
Blood samples for blood gases were taken from a previously injected plastic injector with heparin
(500 IU of liquid heparin for 1 ml of blood), and after blood was taken, the injector was sterilized by air
bending and the measurements were made within 15 minutes. In the collected blood samples, pH, partial
carbon dioxide (PCO2), partial oxygen pressure (PO2), total carbon dioxide concentration (TCO2), base
saline (BE), bicarbonate (HCO3
-), chlorine (Cl-), sodium (Na+), potassium, calcium (Ca++) were evaluated
by portable blood gas analyzer (Epoc Portable Vet) using commercial cards.
Statistical Analyses
Descriptive statistics (distribution, mean, standard deviation, standard error, etc.) relating to measurements
made primarily were included in the study. However, one-way ANOVA and Duncan test were
used to test differences between groups. In addition, Repeated Measures ANOVA was used to compare
repeated measures at 1, 3, and 7 hours for the same subjects. The level of significance was determined
as 0.05 in the analyses made in the study and the SPSS 18.0 for Windows package program was used in
the analysis of the data obtained.

RESULTS

Clinical, hematological, biochemical and immunologic findings of the animals in the study and control
group are presented below.
Clinical Findings
When the heart and respiratory frequencies (P and R / min) were compared with the other groups, the highest
averages were observed in the pre-study control and study group (p <0.05), the P and R averages obtained at 1 (p
<0.05). Clinical findings such as skin elasticity, clinical presentation of dehydration, severity, and frequency of
diarrhea and absorption reflexes were observed to respond well to treatment as time progressed (Table 1).
Hematological Findings
When the WBC levels were compared before the study, there was a statistically significant difference
between the study and control groups (p< 0.05). The first and 7th-hour study groups had the lowest average
of WBC, although no significant differences were seen in terms of statistics (p>0.05) between the
study and control groups at the same hours. RBC and HTC showed a gradual decline in their median and
fell to the lowest level at 7 hours after the study (p <0.05). In opposition, the highest levels of MCHC
level averages were obtained in the 7th hour post-study. MCV averages were found to be lower than the
control group mean (p <0.05), but significantly higher than the mean 3rd and 7th hours (p <0.05). There
was no statistically significant difference between the groups before and after the study in terms of HBG
and MCH level averages (p> 0.05).
Serum Biochemical Findings
The AST and GGT levels at 1, 3, and 7 hours after the study did not show any statistically significant differences
between the control and study groups at the same time points (p> 0.05), and the AST and GGT levels
of the control and study groups and the difference between the groups was statistically significant (p <0.05).
After the study, it was determined that the mean UREA and CREA decreased gradually in the later
time periods and there was a statistically significant difference (p <0.05) between the averages in terms
of time periods.
When examined in terms of groups after the study, the TP and ALB levels increased gradually with
time and this increase was statistically significant (p <0.05), but there were no statistically significant
differences (p> 0.05) between the time period and study groups.
The most interesting change was found in the GLU levels. On days 1, 3, and 7 after the study, both
control and study groups had a gradual increase in GLU levels over time, and this increase was statistically
significant in terms of timescales. As a matter of fact, significant differences were observed statistically between
the control and study groups’ GLU averages at the 1st and 3rd hours after the study (p <0.05) and it
was observed that the control group levels were higher than those of the study group at this time period.
Blood Gas Analyses Findings
It was determined that the mean values of pH, PCO2 and TCO2 were found to be statistically significant
(p <0.05) in both groups at the 1st, 3rd and 7th hours in terms of both study and control groups when
compared to pre-study. Interestingly, pO2 levels were within normal limits in all groups, but pre-study
high levels decreased significantly (p <0.05) from statistically significant (p <0.05), and there was no
significant difference between groups at each time point (p> 0.05).
The most striking finding is that there were found to be significant differences in the HCO3
- levels
of the control groups after each study period (bicarbonate given). However, this statistic was noticed
at the 7th hour of the difference and there was no difference between the control and the study groups’
HCO3
- levels at the 7th hour.
It was found that the highest LACT levels did not show any significant difference in terms of pre-study
control and study groups (p> 0.05), the control group showed similarity to the 7th hour average, and
the mean of all other time period control groups was lower than the LACT mean of the study groups).
It was determined that the K+ levels showed a gradual decrease with time and this decrease was faster
and statistically significant (p <0.05) than in the study group (p <0.05). Contrary to this situation, the
highest Na+ level averages were obtained in the control group at the 3rd and 7th hours. When compared
to the other groups, this difference was statistically significant (p <0.05). Chlor levels were found to be
faster in the study group (NaCl-given group) than in the control group at all time intervals, and this difference
was statistically significant (p <0.05). It was also found that each time progression of Ca++ levels
made a difference between the study groups, that these reached the highest level at the 7th hour and that
the differences were statistically significant (p <0.05) (Table 5).

DISCUSSIONS

Respiratory problems which lead to reduce oxygen-binding capacity of the hemoglobin are frequently
seen in respiratory acidosis (Greenbaum 2004). Clinical respiratory problems and the high respiratory
frequencies obtained in this study can be considered as a sign of this. Özcan and Akgül (2004) reported similar
findings and reported that clinical symptoms of cold, moderate and severe dehydrated diarrhea were
abundant in calves given 8.4% NaHCO3, 0.9% NaCl- and 5% dextrose in appropriate amounts and times.
It has been reported that dehydration-related increases in blood parameters such as HTC and HB in diarrheal
calves have been observed to decrease significantly after fluid therapy (Özcan, C., Akgül, Y. 2004).
It was determined that the pre-study high levels of WBC, RBC, HTC, MCV measurements in this study
showed a gradual decrease with the onset of the study, as the high levels were initially higher due to diar122
rhea-induced dehydration. In our study, there was no difference between the groups in terms of HB levels
and it was seen that the hematological findings including the HB level obtained were in agreement with
the findings reported by Öcal et al. (2006). In our study, although there were significant differences in HTC
levels between the two groups at different time periods, no significant difference was observed between LR
and SF groups at the same time. Similar findings have been reported by Martini et al. (2013).
In this study, when compared with pre-treatment groups, AST, GGT, and UREA and CREA concentrations
were gradually decreased in terms of time intervals in both control and study groups with the
start of treatment. Conversely, however, TP and ALB concentrations, which were low for the pre-study,
gradually increased in the later time periods in both groups after the study started. The most interesting
change was in GLU levels. The highest GLU level was observed in the 7th-day control group, and the
lowest GLU levels were measured in the pre-study control and study group. In all time periods, the
GLU levels were higher in the control group than in the study group, and this is thought to be due to the
metabolism of lactate in the fluid given in the control group to GLU.
Abdalmalek (1987) reports that TP and ALB levels decrease in Escherichia coli, corona, and rotavirusinfected
diarrhea while the levels of urea decrease, but that the levels of GLU increase. In our study, low
levels of GLU, unlike those reported by the researcher, could be attributed to lack of animal ingestion and
insufficiency of lactate metabolism. As a matter of fact, it is reported that a loss of normocyclic anion develops
in cases of diarrhea and lactic acidosis is always present in cases and high blood LACT level as it can not
be detected (Cieza, J.A. et al. 2013). A higher blood serum BUN/creatinine ratio than 20/1 indicates prerenal
azotemia and lack of perfusion (Hanna, J.D. et al. 1995). The fact that the ratio of UREA and CREA measured
in this study is high and that there is a presence of metabolic acidosis is a sign of a problem in kidney buffer
mechanisms. In our study, we found a gradual increase in TP and ALB levels at baseline, which was low at the
beginning, and the results we obtained showed that TP and ALB levels in diarrheal calves were high before
treatment and this was consistent with what is reported by many researchers (Kiowa et al. 1990). It differs
from Özcan and Akgül (2004), who reported that the high level was due to hemoconcentration.
When the blood pH was compared to the post-study, it was found that it was significantly lower
before the study, that it was rapidly normalized after treatment, and that no significant difference was
observed between the effects of PS and LRs on pH. These findings are consistent with findings reported
in a similar study (Martini, W.Z. et al. 2013).
It has been reported that low TCO2 levels are indicative of metabolic or respiratory acidosis, the differential
can be determined by the change in HCO3
- concentration, and the low concentration of HCO3
-can only indicate metabolic acidosis (Narins and Gardner 1981). In our study, the decrease in TCO2
levels, accompanied by HCO3
- levels, proves that the case is metabolic acidosis. We also found that the
findings support the finding that LRs was more effective than PS in normalizing serum HCO3
- levels
(Martini, W.Z. et al. 2013) when compared with the findings of PS.
Metabolic acidosis develops in diarrheal frogs as a response to intracellular K+ extracellular space,
extracellular H+, Na+, and Cl- into the intracellular space. As a result of these transitions, blood K+ level
increases and Na+ level decreases (Lewis, L.D., Phillips, RW. 1978). In our study, high K+, low Na+
and Cl- levels in pre-treatment groups in diarrheal calves were consistent with those reported by the
researcher (Özcan, C., Akgül, Y. 2004; Radostits, O.M. et al. 2006).
Serum physiologic and LRs are both crystalloid fluids and are widely used for the reconstitution of
liquid electrolyte balance (Maier, R.V. 1997). However, it has been reported that LRs, which has better
effects on the heart and acid-base balance and oxygen excretion, is more compatible with body fluids
(Martini, W.Z. et al. 2013) when compared to NaCl with high Na+ and Cl- levels and 5.0 pH. In our study,
it was also observed that the LR solutions did not make a difference according to NaCl- in terms of K+
levels, the Na+ and Cl- levels were higher in the NaCl- given group, but the LRs was more effective in the
healing the general rehydration and electrolyte balance. Unlike our study, Mahajan et al. (2012) reported
that there was no significant difference in the healing effects of PS and LRs on the blood pH in children
with acute diarrhea.
Consequently, LRs use resulted in a better clinical response than NaCl- illustrated by a more rapid and more
appropriate physiological correction and calves with metabolic acidosis were cured using this treatment.

REFERENCES

1. ABDELMALEK, B. (1987). Total protein, urea and blood sugar in healthy calves and calves with diarrhea
[in Bulgarian]. In: Veterinarno-medicinski nauki, vol. 24(4), pp. 55-58. ISSN 0506-8215.
2. CHO, Y.I., HAN, J.I., WANG, C., COOPER, V., SCHWARTZ, K., ENGELKEN, T., YOON, K.J. (2013). Casecontrol
study of microbiological etiology associated with calf diarrhea. In: Vet. Microbiol, vol. 166(3-4), pp. 375-
385. DOI 10.1016/j.vetmic.2013.07.001
3. CIEZA, J.A., HINOSTROZA, J., HUAPAYA, J.A., LEON, C.P. (2013). Sodium chloride 0.9% versus
Lactated Ringer in the management of severely dehydrated patients with choleriform diarrhoea. In: Journal of
Infection in Developing Countries, vol. 7(7), pp. 528-532. DOI 10.3855/jidc.2531
4. EMIRALIOĞLU, M., ÖZÇELIK, U. (2014). Hipokalsemi ve oksijen tedavisi. In: Çocuk Sağlığı ve
Hastalıkları Derg, vol. 57(1), pp. 50-60. ISSN 0010-0161.
5. FOSTER, D.M., SMITH, G.W. (2009). Pathophysiology of diarrhea in calves. In: Veterinary Clinics of
North America: Food Animal Practice, vol. 25, pp. 13-36. DOI 10.1016/ j.cvfa.2008.10.013
6. GREENBAUM, L.A. (2004). Pathophysiology of body fluids and fluid therapy. In: BEHRMAN, R.E., KLIEGMAN,
R.M., JENSON, H.B., eds. Nelson Textook of Pediatrics. 17th ed., pp. 223-234. ISBN 978-0721695563.
7. HALL, G.A., JONES, P.W., MORGAN, J.H. (1992). Calf Diarrhoea. In: Andrews, A.H. et al., ed. Bovine
Medicine: diseases and husbandry of cattle. USA: Blackwell Science, pp. 154-180. ISBN 0632030399.
8. HANNA, J.D., SCHEINMAN, J.I., CHAN, J.C.M. (1995). The kidney in acid-base balance. In: Pediatric
Clinics of North America, vol. 42(6), pp. 1365-95. DOI 10.1016/S0031-3955(16)40089-1
9. HUNT, E. (1992). Diarrheal diseases of neonatal rurninants. In: Howard, J.L., ed. Current Veterinary Therapy:
Food Animal Practice. 3th ed. Philadelphia: Saunders, pp. 103-110. ISBN 978-0721636337.
10. KEE, J.L., PAULANKA, B.J, POLEK, C. (2010). Handbook of fluid, electrolytes, and acid-base imbalances.
3rd ed. Delmar: Cengage Learning, USA, pp. 127-152. ISBN 978-1435453685.
11. LEWIS, L.D., PHILLIPS, R.W. (1978). Pathophysioiogic Changes due to coronavirus-induced diarrhea in
the calf. In: Journal of the American Veterinary Medical Association, vol. 173(5), pp. 636-642. ISSN 0003-1488.
12. LORENZ, I., KLEE, W. (2007). Neonatal calf diarrhoea -something old, something new. In: Cattle Practice,
vol. 15, pp. 146-151. ISSN 0969-1251.
13. MAGANCK, V., HOFLACK, G., OPSOMER, G. (2014). Advances in prevention and therapy of neonatal
dairy calf diarrhoea). a systematical review with emphasis on colostrum management and fluid therapy. In: Acta
Veterinaria Scandinavica, vol. 56(1), pp. 75-83. DOI 10.1186/s13028-014-0075-x
14. MAHAJAN, V, SAJAN, S.S., SHARMA, A., KAUR, J. (2012). Ringer’s lactate us Normal saline for children
with acute diarrhea and severe dehydration: a double blind randomized controlled trial. In: Indian Pediatrics,
vol. 49(12), pp. 963-968. ISSN 0019-6061.
15. MAIER, R.V. (1997). Shock. In: GREENFUELD, L.J. et al., eds. Surgery: Scientific Principles and Practice.
2nd ed. Philadelphia: Lippincott-Raven, pp. 182-215.
16. MARTINI, W.Z., CORTEZ, D.S., DUBICK, M.A. (2013). Comparisons of normal saline and lactated Ringer’s
resuscitation on hemodynamics, metabolic responses, and coagulation in pigs after severe hemorrhagic shock. In:
Scandinavian Journal of Trauma, Resuscitation and Emergency Medicine, vol. 21, pp. 86-95. ISSN 1757-7241.
17. MULLINS, R.J. (1996). Management of Shock. Stamford, Appleton &Lange, Philadelphia, USA.
18. NARINS, R.G., GARDNER, L.B. 1981). Simple acid-base disturbances. In: Medical Clinics of North America,
vol. 65(2), pp. 321-346. ISSN 0025-7125.
19. ÖCAL, N., DURU, S.Y., BAĞCI, B.B., GAZYAĞCI, S. (2006). İshalli buzağılarda asit-baz dengesi
bozukluklarının saha şartlarında tanı ve sağaltımı. In: Kafkas Universitesi Veteriner Fakultesi Dergisi, vol. 12(2),
pp. 175-183. ISSN 1300-6045.
20. ÖZCAN, C., AKGÜL, Y. (2004). Neonatal İshalli Buzagılarda Hematolojik, Biyokimyasal ve Elektrokardiyografik
Bulgular. In: Yuzuncu Yil Universitesi Veteriner Fakultesi, vol. 15(1-2), pp. 123-129.
21. RADOSTITS, O.M., GAY, C.C., HINCHCLIFF, K.W., CONSTABLE, P.D., eds. (2006). Veterinary Medicine.
A textbook of the diseases of cattle, sheep, goat, pigs and horses. 10th ed. BailliereTindall, London, 2065 p.
ISBN 9780702039911.
22. ROUSSEL, A.J. (1993). Fluid Therapy, Transfusion, and Shock Therapy. In: Howard, J.L., ed. Current
Veterinary Therapy: Food Animal Practice. 3th ed. Philadelphia: Saunders, 8 p. ISBN 978-0721636337.
23. SARNAIK, A.P., HEIDEMANN, S.M. (2007). Respiratory pathophysiology and regulation. In: BEHRMAN,
R.E., KLIEGMAN, R.M., JENSON, H.B., eds. Nelson Textbook of Pediatrics 18th edit. Philadelphia:
Saunders Company, pp. 1719-1726. ISBN 9781437721805.