THE EFFECTS OF ORALLY GIVEN HIGH RATE CARBOHYDRATE
ON SOME PATHOGENS THAT PLAY AN IMPORTANT ROLE IN
ETIOLOGY OF THE DIARRHEA IN CALVES
Bulent ELITOK
Afyon Kocatepe University, Faculty of Veterinary Medicine, Department of Internal Medicine,
03200-Afyonkarahisar/Turkey, E-mail: elitok1969@hotmail.com
Abstract. This study was carried out on 30 adult calves which completed their rumen development aged from
three to six months old in the public held. Twenty calves received totally 1500 ml of propylene glycol per day 3-5
times/day, assigned as study group, while 10 calves that were found to be clinically healthy and didn’t receive any
additional treatments served as control group. Clinical, stool, hematological and blood biochemical examinations
were performed in all the animals. At the end of the study; it had been found that propylene glycol caused mild diarrhea
(15%) and some slightly respiratory system problems (10%), decreased the number of calves which showed
fecal-pathogenic agents in their feces, and didn’t cause significant problems in the liver. Consequently; it was determined
that a glucose precursor propylene glycol could be used safely in adult calves, and it might help to reduce
fecal contamination to neonates by decreasing the number of fecal pathogens, besides increased productivity.
Key words: Calves; Propylene glycol; Pathogenic agents; Faeces.
INTRODUCTION
Calf diarrhea is still the most frequent and significant economic loss in cattle breeding, despite the big
improvements in herd management, housing conditions, care, nutrition and biopharmaceuticals (Izzo,
M.M. et al. 2011). These studies report that while calf deaths in the neonatal period are higher than in
the adult turnover, adult deaths are also at a significant level.
Despite the large number of studies carried out at early ages, the health of older adults calves is less
researched. Studies of morbidity have shown that the diarrhea and respiratory diseases are the most important
disease groups seen in the older calves (Perez, E. et al. 1990; Olsson, S.O. et al. 1993; Svensson, C. et
al. 2003). As a matter of fact, the incidence of diarrhea is decreasing with age (Frank, N.A., Kaneene, J.B.
1993; Bendali, F. et al. 1999), while the risk of developing diarrhea in the first months of life is low.
Propylene glycol (PG) is one of the most widely used substances for energy supply in cattle, decrease
in the amount of ketone bodies, increase in yield and elimination of losses during disease (Gordon, J.L.
et al., 2017; Raboisson, D. et al. 2014; Gohary, K. et al. 2016; Bjerre-Harpoth, V. et al. 2016). The use of
high doses of PG may lead to diarrhea as well as toxic effects (Fiume, M.M. et al. 2012). Sabbioni, A. et
al. 1999) reported that long-term administration of PG (50 ml/animal/ day) in the high dose was caused
to slightly toxic effects in the liver as well as diarrhea formation.
The aim of this study was to investigate the effects of higher dose of PG on the presence of pathogenic
agents (virus, bacteria, protozoa) in the feces as well as diarrhea formation in calves in their older ages.
MATERIALS AND METHODS
Animal Material:
The study was carried out on 30 calves who developed rumen, three to six months old in public held.
Twenty calves were recieved 1500 ml of propylene glycol per day 3-5 times/day in total, assigned as
the study group (ÇG), while 10 of the calves were found to be clinically healthy and without additional
treatment served as control (KG).
The present study was carried out within the framework of ethical rules of the Ethical Committee of Animal
Experiments of Afyon Kocatepe University with the reference number of AKUHADYEK 197-17, 17.
CAREER. 69 and the Afyon Kocatepe University Scientific Research Projects Coordination Unit (BAPK).
Clinical Examinations:
Body temperature, respiration and heart rates, ruminal contractions in 5 minutes along with diarrhea, dehydration
and appetite control were determined using the methods described by Blood and Radostits, (1989).
Hematological Examination:
Hematological parameters such as total leukocyte (WBC), erythrocyte (RBC), mean corpuscular voluŞtiinţa
me (MCV), hematocrit (HCT), mean cell hemoglobin (MCH), mean corpuscular hemoglobin concentration
(MCHC), hemoglobin were measured by Hemocellcounter (Mindray Hemocell Veterinary Model).
Blood Biochemical Tests:
Some blood parameters such as aspartate aminotransferase (AST), gamma-glutamyltransferase
(GGT), glucose (GLU), albumin (ALB) and total protein (TP were measured on Roche Cobas C111
model autoanalyzer using commercial kits.
Viral and Parasitological Experiments:
The commercial Rapid Test Kit (Quatro Vet Uni-Strip Kit, C-1540, Coris BioConcept, Belgium) was
used to verify rotavirus, coronavirus, cryptosporidium, giardia and E. coli in fecal samples. The presence
of Eimeria spp. in the stool was made using the native examination method of fresh faeces (Blood
and Radostits, 1989).
Statistical Analyzes:
One-way ANOVA and Duncan’s test were used to test differences between groups. Also, Repeated
Measures ANOVA was used for repeated measures in comparing the measurements at different times for
the same individuals. The level of significance was determined as 0.05 in the analyses made in the study
and SPSS 18.0 for Windows package program was used in the analysis of the obtained data.
Results
Clinical, hematological and biochemical examination findings with the presence of pathogens in the
faeces of animals in the study and control group are shown as below.
Existence of Patients in the Stool and Clinical Examination Findings:
Findings related to the presence of pathogens, dehydration, diarrhea and loss of appetite before and
after the application of propylene glycol (PG) in the study group (SG) animals are shown in Graphic 1,
in relation to the parameters mentioned in the Control Groups (CG) the data are shown in Graphic 2.
The animals in the SD received a gradual increase in the level of diarrhea following the administration
of PG and the highest number (15% of the animals) on day 5, the last day of the study. Similarly, dehydration
and loss of appetite reached 5% of all animal populations (n= 20) on day 5, although observed in
different animals. From the point of view of CG animals, no change was found in terms of the parameters
mentioned. Interestingly, there were detected in 2 animals (10% of the entire polution) of the animals (n =
20), coronavirus, cryptosporidium and E. coli, and 3 (15%) Eimeria chart 1) in SG. On the days following
PG administration, a reduction in fecal and efficacious animal numbers was observed, with the lowest
numerical value being determined on the fifth day of the last day of administration. Compared to the preadministration
effluent, only 5% of the total animals (n = 1) of pathogenic microorganisms were found to
have reduced half-life (50%) on the 5th day after the administration of PG, and the route and corona virus,
cryptosporidium and E. coli pathogens 1 animal. Similarly, the number of animals found in Eimeria in their
stools decreased by 30% on the 5th day.,10% of total animals), cryptosporidium (20% of total animals), E.
coli (20% of total animals) and Eimeria (10% of total animals) (20% of total animals), 3 animals (corresponding
to 33% of the total animals) were found in the mix (Graph-2). There was no numerical change in
the duration of the study from the point of the CG animals in terms of smearing of excreta.
The clinical parameters measured in CG and SG animals are shown in Table 1.
When Table 1 is examined; SG animals were observed to be significantly higher (p <0.05) in the
statistically significant (p <0.05) level of pre-administration and body temperature of the CG animals,
within the normal limits, following days of PG administration. However, no statistically significant
difference was found between recalcitrant days of CG animals and the average of the animals before
PG administration (p> 0.05). There was no statistically significant difference between consecutive days
averages of SC animals after PG administration (p> 0.05). A similar statistic was also found in terms of
the given heart and respiratory frequencies. Although the respiratory and cardiac frequencies of all CG
and SG animals were within normal limits, there were no statistically significant changes between the
QoL intervals and the pre-PG averages of PG animals (p> 0.05) (p> 0.05), but the PG administration
was significantly higher (p <0.05) in the statistically significant difference between the respiratory and
cardiac frequency averages before and after the administration of the CG animals, within the normal
range on the following days. In addition, 2 animals (10%) were found to have a problem of respiratory
air, australic lung sounds hardened, mild respiratory system problem. Although it led to a slight increase
in the number of rumen contractions of the PG for 5 minutes, it was still statistically insufficient to make
a statistically significant difference (p> 0.05) in terms of the rumen contractions at 5 min during the
study period in CG and SG animals.
Hematological Examination Findings:
Avarage rates of hematological findings observed in this study are shown at Table 2.
When Table 2 is examined; it was determined that the highest WBC, RBC, HB and HCT averages
(7.90 ± 2.10, 8.60 ± 3.00, 9.90 ± 2.40, 30.60 ± 4.20) were obtained on the 5th day after PG administration.
When the comparisons between the groups were examined, it was understood that CG did not show
any difference between the mean values determined for the parameters mentioned in all the time periods
of animals and the mean of the pre-PG data before the application (p> 0.05). The mean values obtained
after the PG administration in the SG animals were significantly higher than the mean values of the PG
animals before and after the PG administration in terms of statistical significance (p> 0.05), although
the mean values of the SG animals in terms of the stated parameters were not statistically significant
(p> 0.05). In terms of MCV, MCH and MCHC levels measured in this study, there was no statistically
significant change in terms of time intervals between groups (p> 0.05).
Metabolic Profile Findings:
The averages of the data obtained from blood biochemical specimens in this study are shown in Table 3.
According to this Table the highest average values of AST and GGT enzyme levels were formed on
the 5th day after PG administration (86.50 ± 15.30, 296.70 ± 68.30, respectively) and gradually increased
from day 1 to day 5 after PG administration, (p <0.05) was found to be statistically significant.
Similarly, it was found that the mean values of the SG animals were significantly higher (p <0.05) in
terms of statistical significance than the mean of CG before PG and PG, but there was a statistically significant
difference between the mean values of PG and PG pre-PG averages (p> 0.05). Similar, but more
pronounced, elevation measurements have also been observed in terms of GLU concentrations. There
was no statistically significant difference (p> 0.05) in terms of the GLU concentration averages of SG
animals before CG and PG administration (p> 0.05) and the mean values obtained at all time intervals
of CG were higher than the average of SG animals before PG administration. Similar elevations were
also observed in EC animals after the PG administration and the difference between the average of GLU
concentration levels in the days following PG administration was statistically significant (p <0.05), the
highest level was found on the 5th day after PG administration (98.47 ± 7.26). In terms of TP and
ALB concentration levels, there was no significant difference between both groups (p> 0.05).
Discussion
In our literature reviews, we unfortunately did not find a lot of works that directly addressed the
effects of PG on diarrhea in adult icebergs and possible pathogens that are spread by feces in this diarrhea.
However, PG, a carbohydrate percursor, is known to cause diarrhea by altering osmotic pressure in
the digestive tract (Hammer et al., 1989; Hendrickson, 2017; Trabue et al., 2007). In our present study,
it was determined that diarrhea developed in 3 (15%) of the ED animals at the 3rd day following PG
administration, whereas no diarrhea was detected before the first mailling of the CG animals and before
the PG administration of the SG animals. which can lead to the formation of the nature.
As it is well known, even if pathogenic agents do not form the disease table on adult calves, they
continue to be found in facultative form in the digestive tract flora in healthy calves (Janke, B.H. 1989;
Fagan, J.G. et al. 1995; Uhde, F.L. et al. 2008). In our work a total of 1 animal rota and corona virus were
found in CG, 2 animals Eimeria, 2 E. coli and 2 cryptosporidium were detected. In SG animals, after 2
weeks of PG administration, a total of 2 animal rota virus, 1 animal corona virus, 2 animals Eimerai, 2
cryptosporidium and 3 E. coli were found to be in agreement with the researchers.
Recently, PG has been the most carefully studied glucogenic supplements, and some researchers
(Robinson, E. and Sprayberry, K. 2009) mention that antibacterial and antifungal effects may be present.
T.O. Thorgeirsdottir et al. (2003) reported that PG increased antiviral efficacy in combination with antiviral
drugs at different concentrations. T.M. Nalawade et al. (2015) claimed that PG had bacteriocidal
effects on many bacteria, especially E. coli. M. Khaw et al. (1995) reported an increase in the activity
of antiprotozoal drugs used in combination with PG. As a matter of fact, the decrease in the number of
animals showing these factors in the feces in the days after the PG application in our current study, espeŞtiinţa
cially on the 5th day, especially on the last day, supports the finds of the researchers even if the number
is small.
As a result of PG administration at high doses; (Nielsen, N.I. et al. 2004; McClanahan, S. et al. 1998;
Ivany, J.M. and Anderson, D.E. 2001), as well as respiratory system-related disorders such as depression,
ataxia, excessive salivation, malodorous respiration, and malodorous fecal symptoms. As a matter
of fact, although in our study, we could not mention a numerical calf population enough to support this
data following the PG application, it was found that all the factors were detected at the same time, 1 of
the animals in the SG had a bad smell of respiratory air and 2 animals had a slight garlic-like odor its
appearance, supports the above-mentioned views.
The present study supports the findings of researchers (Munday, R. and Manns, E. 1994) who reported
that the lowest levels of HGB and RBC, as well as the mean levels of HGB and RBC, were detected
on the 5th day of the SG and that oxidative stress-related hemolytic anemia could be formed in animals
given PG-like substances.
PG, a glucose precursor, is often used to relieve energy needs. Frequent use of PG does not produce
toxic effects (Fiume, M.M. et al. 2012; Ivany, J.M. and Anderson, D.E. 2001; DeFrain, J.M. et al. 2014),
it can also cause diarrhea. Along with the diarrhea table; hypovolemia (dehydration and loss of metabolites),
metabolic acidosis, hyperkalemia, renal insufficiency (Cho, Y. and Yoon, K. 2014; Steven et
al. 2007). The higher levels of GLU levels measured in our study compared to the KG average after PG
administration support the findings of these investigators. A. Sabbioni et al. (1999) reported that blood
triglycerides and NEFA decreased, while long-term use improved mild toxicity in the liver, while providing
PG (50 ml/animal/day) carcass increase in adult icebergs. In our study, PG was applied for 5 days
and the AST measured in the icebergs supports the GGT enzyme levels reported by these researchers,
which are higher than the average of CG, while staying within normal limits. Emery et al.,1964), on
the other hand, argue that even with high doses such as 2000 g/cow/day, PG does not produce any side
effects or toxicity data. In our study, the measurement of blood results from biochemical investigations
is closer to normal than that reported by this investigator.
Consequently; PG, a commonly used carbohydrate precursor, causes little diarrhea as a result of a
5-day trial, causing some increases in some of the liver’s enzymes, within normal limits, but these elevations
are not enough to claim a toxicity. In addition, it has been concluded that PG causes a numerical
decrease in fecal and efficacious animal numbers and that it would be useful to use it on adult icebergs.
REFERENCES
1. BARTELS, C.J.M., HOLZHAUER, M., JORRITSMA, R., SWART, W.A.J.M., LAM, T.J.G.M. (2010).
Prevalence, prediction and risk factors of enteropathogens in normal and non-normal faeces of young Dutch dairy
calves. In: Preventive Veterinary Medicine, vol. 93(2-3), pp. 162- 169. ISSN 0167-5877.
2. BENDALI, F., BICHET, H., SCHELCHER, F., SANAA, M. (1999). Pattern of diarrhoea in newborn beef
calves in south-west France. In: Veterinary Research, vol. 30(1), pp. 6-74. ISSN 1993-5412.
3. BJERRE-HARPOTH, V., STORM, A.C., VESTERGAARD, M., LARSEN, M., LARSEN, T. (2016).
Effect of postpartum propylene glycol allocation to over-conditioned Holstein cows on concentrations of milk
metabolites. In: Journal of Dairy Research, vol. 83(2), pp. 156-64. ISSN 1469-7629.
4. CHO, YI., YOON, KJ. (2014). An overview of calf diarrhea - infectious etiology, diagnosis, and intervention.
In: Journal of Veterinary Science, vol. 15(1), pp. 1-17. DOI 10.4142/jvs.2014.15.1.1
5. DEFRAIN, J.M., HIPPEN, A.R., KALSCHEUR, K.F., JARDON, P.W. (2004). Feeding glycerol to transition
dairy cows, Effects on blood metabolites and lactation performance. In: Journal of Dairy Science, vol. 87, pp.
4195-4206. ISSN 0022-0302.
6. EMERY, R.S., BURG, N., BROWN, L.D., BLANK, G.N. (1964). Detection, occurrence and prophylactic
treatment of borderline ketosis with propylene glycol feeding. In: Journal of Dairy Science, vol. 47, pp. 1074-
1079. ISSN 0022-0302.
7. FAGAN, J.G., DWYER, P.J., QUINLAN, J.G. (1995). Factors that may affect the occurrence of enteropathogens
in the feces of diarrhoeic calves in Ireland. In: Irish Veterinary Journal, vol. 48, pp. 17-21. ISSN 2046-0481.
8. FIUME, M.M., BERGFELD, W.F., BELSITO, D.V., HILL, R.A., KLAASSEN, C.D., LIEBLER, D.,
MARKS, J,G., SHANK, R.C., SLAGA, T.J., SNYDER, P.W., ANDERSEN, F.A. (2012). Safety assessment of
propylene glycol, tripropylene glycol, and PPGs as used in cosmetics. In: International Journal of Toxicology, vol.
31(5), pp. 245-260. ISSN 1091-5818.
9. FRANK, N.A., KANEENE, J.B. (1993). Management risk factors associated with calf diarrhea in Michigan
dairy herds. In: Journal of Dairy Science, vol. 76, pp. 1313-1323. ISSN 0022-0302.
10. GOHARY, K., OVERTON, M.W., Von MASSOW, M., LEBLANC, S.J., LISSEMORE, K.D., DUFFIELD,
T.F. (2016). Economic value of ionophores and propylene glycol to prevent disease and treat ketosis in Canada. In:
Canadian veterinary journal, vol. 57(7), pp. 733- 740. ISSN 0008-5286.
11. GORDON, J.L., LEBLANC, S.J., KELTON, D.F., HERDT, T.H., NEUDER, L., DUFFIELD, T.F. (2017).
Randomized clinical field trial on the effects of butaphosphan-cyanocobalamin and propylene glycol on ketosis
resolution and milk production. In: Journal of Dairy Science, vol. 100(5), pp. 3912-3921. ISSN 0022-0302.
12. HAMMER, H.F., SANTA ANA, C.A., SCHILLER, L.R., FORDTRAN, J.S. (1989). Studies of
osmotic diarrhea induced in normal subjects by ingestion of polyethylene glycol and lactulose. In: Journal of
Clinical Investigation, vol. 84(4), pp. 1056-1062. ISSN 0021-9738.
13. HENDRICKSON, K. (2017). Properties of Propylene Glycol [accessed: 30 Aug. 2017]. Avalable: www.
livestrong.com/article/116991-properties-propylene-glycol/
14. IVANY, J.M., ANDERSON, D.E. (2001). Propylene glycol toxicosis in a llama. In: Journal of the American
Veterinary Medical Association, vol. 218, pp. 243-244. ISSN 0003-1488.
15. IZZO, M.M., KIRKLAND, P.D., MOHLER, V.L., PERKINS, N.R., GUNN, A.A., HOUSE, J.K. (2011).
Prevalence of major enteric pathogens in Australian dairy calves with diarrhoea. In: Australian Veterinary Journal,
vol. 89, pp. 167-173. DOI 10.1111/j.1751-0813.2011.00692.x.
16. JANKE, B.H. (1989). Protecting calves from diarrhea. In: Veterinary Medicine, vol. 84, pp. 803- 811.
17. KHAW, M., CLAIRE, B., PANOSIAN, C.B. (1995). Human Antiprotozoal Therapy, Past, Present, and
Future. In: Clinical Microbiology Reviews, vol. 8(3), pp. 427-439. ISSN 0893-8512.
18. McCLANAHAN, S., HUNTER, J., MURPHY, M., VALBERG, S. (1998). Propylene glycol toxicosis in a
mare. In: Veterinary and Human Toxicology, vol. 40, pp. 294-296. ISSN 0145-6296.
19. MUNDAY, R., MANNS, E. (1994). Comparative toxicity of prop(en)yl disulfides derived from Alliaceae,
Possible involvement of 1-propenyl disulfides in onion-induced hemolytic anemia. In: Journal of Agricultural and
Food Chemistry, vol. 42(4), pp. 959-962. DOI 10.1021/jf00040a023
20. NALAWADE, T.M., BHAT, K., SOGI, S.H.P. (2015). Bactericidal activity of propylene glycol, glycerine,
polyethylene glycol 400, and polyethylene glycol 1000 against selected microorganisms. In: Journal of International
Society of Preventive and Community Dentistry, vol. 5(2), pp. 114-119. DOI 10.4103/2231-0762.155736.
21. NIELSEN, N.I., INGVARTSEN, K.L. (2004). Propylene glycol for dairy cows. A review of the metabolism
of propylene glycol and its effects on physiological parameters, feed intake, milk production and risk of ketosis. In:
Animal Feed Science and Technology, vol. 115(3-4), pp. 191- 213. DOI 10.1016/j.anifeedsci.2004.03.008
22. OLSSON, S.O., VIRING, S., EMANUELSON, U., JACOBSSON, S.O. (1993). Calf diseases and
mortality in Swedish dairy herds. In: Acta veterinaria Scandinavica, vol. 34(3), pp. 263-269. ISSN 0044-605X.
23. PEREZ, E., NOORDHUIZEN, J.P.T.M., Van WUIJKHUISE, L.A., STASSEN, E.N. (1990). Management
factors related to calf morbidity and mortality rates. In: Livestock Production Science, vol. 25(1-2), pp. 79-93.
ISSN 0301-6226.
24. RABOISSON, D., MOUNIE, M., MAIGNE, E. (2014). Diseases, reproductive performance, and changes
in milk production associated with subclinical ketosis in dairy cows, a meta-analysis and review. In: Journal of
Dairy Science, vol. 97(12), pp. 7547-7563. ISSN 0022-0302.
25. ROBINSON, E., SPRAYBERRY, K.A. (2009). Current Therapy in Equine Medicine. 6th ed. Elsevier
Health Sciences, USA. 1104 p. ISBN 9781416054757.
26. SABBIONI, A., SUPERCHI, P.BONOMI, A., TAGLIETTI, P. et al. (1999). The use of propylene glycol
in veal calf feeding. In: ASPA Congress, vol. 13, pp. 351-353
27. SVENSSON, C., LUNDBORG, K., EMANUELSON, U., OLSSON, S.O. (2003). Morbidity in Swedish
dairy calves from birth to 90 days of age and individual calf-level risk factors for infectious diseases. In: Preventive
Veterinary Medicine, vol. 58(3-4), pp. 179- 197. ISSN 0167-5877.
28. THORGEIRSDOTTIR, T.O., THORMAR, H., KRISTMUNDSDOTTIR, T. (2003). Effects of polysorbates
on antiviral and antibacterial activity of monoglyceride in pharmaceutical formulations. In: Pharmazie, vol.
58(4), pp. 286-287. ISSN 0031-7144.
29. TRABUE, S., SCOGGIN, K., TJANDRAKUSUMA, S., RASMUSSEN, M.A., REILLY, P.J. (2007).
Ruminal Fermentation of Propylene Glycol and Glycerol. In: Journal of Agricultural and Food Chemistry, vol.
55(17), pp. 7043−7051. DOI 10.1021/jf071076i
30. UHDE, F.L., KAUFMANN, T., SAGER, H., ALBINI, S., ZANONI, R., SCHELUNG, E., MEYLAN,
M. (2008). Prevalence of four enteropathogens in the faeces of young diarrhoeic dairy calves in Switzerland. In:
Veterinary Record, vol. 163(12), pp. 362-366. ISSN 0042-4900.