Abstract
Background
Neonatal calf diarrhea is a major problem in the cattle industry worldwide. Rotavirus and Cryptosporidium parvum are the primary causative agents, especially during the first three weeks of the calf’s life.
Objectives
This study investigated the differences in acid-base, electrolytes, and biochemical parameters of diarrheic calves with infection of either rotavirus or C. parvum.
Methods
A total of 61 Korean native calves (≤ 20 days old) were divided into two groups based on rotavirus or C. parvum infections: rotavirus infection (n = 44) and C. parvum infection (n = 17). The calves with at a specific blood pH range (pH 6.92–7.25) were chosen for comparison. The acid-base, electrolyte, chemistry, and serum proteins were analyzed, Further, fecal examinations were performed.
Results
Compared to C. parvum-infected calves, the rotavirus-infected calves showed lower levels of total carbon dioxide, bicarbonate (HCO3 −), anion gap, total protein, and albumin/globulin ratio, and significantly lower levels of potassium, globulin, and α2-globulin (p < 0.05). The C. parvum-infected calves (r = 0.749) had stronger correlations between pH and HCO3 − than the rotavirus-infected calves (r = 0.598). Compared to rotavirus-infected calves, strong correlations between globulin and α2-globulin, α2-globulin and haptoglobin were identified in C. parvum-infected calves.
Conclusions
This study is the first to investigate acid-base, electrolyte, and biochemical parameters in calves in response to infections of rotavirus and C. parvum. Although rotavirus and C. parvum cause malabsorptive and secretory diarrhea in similar-aged calves, blood parameters were different. This would help establish the diagnostic and treatment strategies.
Keywords: Diarrhea, electrolyte, serum protein, rotavirus, Cryptosporidium parvum
INTRODUCTION
Owing to its high morbidity and mortality, neonatal calf diarrhea continues to be a major problem for the cattle industry worldwide. Calf diarrhea also causes economic loss through the cost of medical treatment, retarded growth, and human labor. Infectious agents and non-infectious factors influence the pathogenesis of calf diarrhea. Various types of virus, bacteria, and protozoa comprise the enteric pathogens of calf. However, rotavirus, coronavirus, Escherichia coli, Salmonella spp., and Cryptosporidium parvum are primary infectious agents in calf diarrhea [1,2,3], and the most frequently identified pathogens in fecal samples are rotavirus and C. parvum [4].
At 1–3 weeks, both rotavirus and C. parvum infections are present in calves [5,6]. Both pathogens induce malabsorptive and secretory diarrhea throughout the small intestine. The mature villous enterocytes are the preferential site for rotavirus and C. parvum invasion, which leads to malabsorption by villous atrophy [7]. However, nonstructural glycoprotein 4 in rotavirus infection plays a major role in secretary diarrhea, while prostaglandin-mediated secretion occurs in C. parvum infection [7,8,9]. In addition, the pronounced sites in the intestine are different. The most severe lesions are the duodenum and proximal jejunum in rotavirus infection, but the distal jejunum and ileum in C. parvum infection [7,8,10,11].
Diarrhea leads to dehydration, acid-base imbalance, and gas and electrolyte disturbances in calves. Dehydration and low levels of pH, Na+, total carbon dioxide (tCO2), bicarbonate (HCO3 −), and base excess (BE) of the extracellular fluid were observed in Korean native (Hanwoo) diarrheic calves [12].
As for total protein, hepatocytes produce plasma proteins including acute phase proteins. B lymphocytes and plasma cells secret γ-globulin. Intestinal malabsorption and malnutrition are associated with hypoalbuminemia [13]. Haptoglobin is a hemoglobin binding α2-globulin and bacteriostatic by preventing microorganisms from utilizing iron [14]. Low levels of total protein and albumin and high α2-globulin levels were present in diarrheic Korean native calves at the 1 to 20 days of age. In addition, Haptoglobin was synthesized more in neonatal diarrheic calves, and not serum amyloid A [15].
To the best of our knowledge, no studies have investigated the differences in acid-base, electrolytes, and biochemical parameters between single natural infections of rotavirus or C. parvum in calves. In this study, we hypothesized that calves would respond to rotavirus or C. parvum infection in different ways because infection mechanisms and pronounced infection regions were different even though both pathogens induce malabsorptive and secretory diarrhea at a similar age. Thus, the objective of this study was to determine the differences in acid-base, electrolyte, and biochemical parameters in rotavirus- or C. parvum-infected diarrheic calves. These findings will help understand the characteristics of diarrhea associated with these pathogens and establish a treatment plan for rotavirus- or C. parvum-infected diarrheic calves.
MATERIALS AND METHODS
Ethics approval and consent to participate
All animal procedures were conducted according to ethical guidelines for the use of animal samples, and were approved by the Jeonbuk National University (Institutional Animal Care and Use Committee Decision No. JBNU 2020-052). All cases were naturally occurring infections. Every diarrheic calf was treated after sampling. All procedures and possible consequences were explained to the managers of the surveyed farm, and written consent was obtained.
Sample collection
The current study used a total of 61 diarrheic Hanwoo calves (native Korean cattle breed), which were of ≤ 20 days old and raised in the Republic of Korea from 2019 to 2020. The calves were housed with their dam and fed colostrum and milk by their dam. Blood samples were collected from the jugular vein of the calves in serum-separation tubes (SST) for serum protein analysis. Acid-base, electrolyte, and chemistry analyses were performed immediately after blood sampling. Fecal samples were collected by experienced veterinarians using digital rectal palpation to induce bowel movements. These fecal samples were stored in 50 mL specimen bottles (SPL Life Sciences, Korea) and transported to the laboratory under refrigeration. The sampling procedures were performed 1–2 days after the calves began to show diarrhea (before the treatment).
Fecal examination
Feces were subdivided into solid, semi-solid, loose, and watery stools. Solid and semi-solid stools were classified as normal while loose and watery stools were classified as diarrhea. Fecal samples were initially screened using a rapid diagnosis antigen test kit (BoviD-5 Ag) to identify antigens associated with pathogens, including C. parvum, Giardia duodenalis, E. coli, coronavirus, and rotavirus. These samples were also examined by reverse transcriptase-polymerase chain reaction targeting bovine viral diarrhea virus (BVDV) and Salmonella spp., as well as the five pathogens noted above. BVDV, coronavirus, rotavirus, E. coli, Salmonella spp., C. parvum, and G. duodenalis were the major pathogens associated with diarrhea in calves [16,17]. To detect Eimeria spp., all fecal samples were suspended in a solution of 2.5% potassium dichromate and then transported to the laboratory. In the laboratory, fecal samples were analyzed to detect oocysts using the floatation methods with Sheather’s solution (saturated sugar solution; specific gravity 1.28) and examined microscopically (×400 magnification) based on the morphological features of Eimeria spp. oocysts.
Experimental group definition
A total of 114 diarrheic calves used in this study had a score over two, based on a previous study [18]. Diarrheic calves (n = 32) that tested positive for at least one of the other pathogens associated with diarrhea, except for rotavirus and C. parvum were excluded; none of these pathogens were detected in fecal samples from clinically healthy calves. Calves (n = 10) with concurrent rotavirus and C. parvum infections were excluded. To avoid the difference of blood acidity while comparing calves with rotavirus to those with C. parvum, we used calves (n = 61) with specific blood pH values (pH 6.92–7.25). The calves were divided into two groups according to the agent: rotavirus infection (n = 44) and C. parvum infection (n = 17). The parameters of clinically healthy calves (n = 57) were used as the reference (Table 1).
Table 1. Descriptive statistics for Hanwoo calves included in the study.
| Variable | Rota (n = 44) | Crypt (n = 17) | p value | Healthy (n = 57) | |
|---|---|---|---|---|---|
| Sex | 0.580 | ||||
| Male | 22 | 7 | 28 | ||
| Female | 22 | 10 | 29 | ||
| Age (day) | 10.64 ± 4.41 | 10.35 ± 3.39 | 0.813 | 11.79 ± 4.32 | |
Data are presented as the mean ± SD.
Rota, diarrheic calves infected with rotavirus; Crypt, diarrheic calves infected with Cryptosporidium parvum; Healthy, clinically healthy calves without diarrhea.
Acid-base, electrolyte, and chemistry analyses
Blood parameters, such as pH, tCO2, partial pressure of carbon dioxide (pCO2), HCO3 −, BE, Na+, K+, Cl−, anion gap (AG), glucose, and blood urea nitrogen (BUN) were measured immediately after obtaining fecal samples using the EC8 + i-STAT cartridge (Abbott, USA). The strong ion difference (SID) was calculated based on the combined electrolyte concentrations as shown below:
| SID = (Na+ + K+) − Cl−. |
Serum protein gel electrophoresis and acute phase proteins
Serum samples were separated from blood by centrifugation. Serum was harvested by centrifuging SST tubes at 3,000 rpm (2,600 g) for 10 min. The serum was frozen and stored at −70°C pending analysis. Serum samples (n = 6) with low quality were excluded (rotavirus group, n = 4; Cryptosporidium group, n = 2). Subsequently, agarose gel electrophoresis was performed to analyse 5 protein fractions (albumin, α1-globulin, α2-globulin, β-globulin, and γ-globulin) using a semi-automated agarose gel electrophoresis system (HYDRASYS 2; Sebia, UK), following the manufacturer’s protocols. Briefly, 30 µL of serum was subjected to the microtechnique assay, electrophoresed for 35 min, stained for 5 min, de-stained for 5 min, and cleared for 30 sec. Excess solution was removed with a glass rod and samples were dried for 10 min and then measured by optical density scanning (HYDRASYS; Sebia). Normal serum was used as a control for measurement accuracy. The results of the serum protein electrophoresis gel were reviewed and interpreted by a laboratory expert.
Analyses for haptoglobin and serum amyloid A were performed using serum samples. Haptoglobin concentrations were assessed using commercial colorimetric kits (Tridelta Development, Ireland) based on haemoglobin binding assay. Serum amyloid A levels were analyzed using sandwich enzyme-linked immunosorbent assay (ELISA) kits (Tridelta Development). The optical densities were read on a microplate reader (BioTek Instruments, USA) at 630 nm for haptoglobin and at 450 nm and 630 nm as a reference for serum amyloid A.
Statistical analyses
Statistical analyses were performed using SPSS software (version 26.0; IBM Corp., USA). The Shapiro-Wilk test and Levene’s test were used for normality analysis and equality of variances for independent t-test. Data are expressed as the mean ± SD. Statistical significance was set at p < 0.05. Pearson’s correlation test was used to determine correlations between the parameters. Pearson’s correlation coefficient is represented as r ≥ 0.7, strong correlation; 0.5 ≤ r < 0.7, moderate correlation; 0.3 ≤ r < 0.5, weak correlation; and r < 0.3, no correlation.
RESULTS
Acid-base, electrolyte, and chemistry analysis
Diarrheic calves infected with rotavirus or C. parvum showed lower levels of pH, tCO2, pCO2, HCO3 −, BE, Na+, SID, and glucose, but higher levels of Cl−, AG, and BUN than clinically healthy calves. The mean levels of tCO2, pCO2, HCO3 −, and BE in blood were different between calves with rotavirus and those with C. parvum, despite similar pH. Calves infected with rotavirus showed lower values of tCO2, pCO2, HCO3 −, and BE than those infected with C. parvum. The levels of Na+, Cl−, and SID were similar between the two diarrheic groups. The difference in AG was over 1.6 mmol/L between calves with rotavirus and C. parvum. Calves with C. parvum showed lower glucose levels than those with rotavirus, but BUN vice versa, even though the difference was insignificant (Table 2).
Table 2. Acid-base, electrolyte, and biochemical analyses of the blood from rotavirus or Cryptosporidium parvum infected calves.
| Parameter | Rota (n = 44) | Crypt (n = 17) | p value | Healthy (n = 57) |
|---|---|---|---|---|
| pH | 7.09 ± 0.08 | 7.09 ± 0.12 | 0.956 | 7.39 ± 0.04 |
| tCO2 (mmol/L) | 11.07 ± 3.94 | 13.12 ± 5.85 | 0.119 | 31.98 ± 3.29 |
| pCO2 (mmHg) | 32.67 ± 10.36 | 36.35 ± 11.58 | 0.233 | 50.24 ± 3.92 |
| HCO3 − (mmol/L) | 10.16 ± 3.73 | 11.81 ± 5.50 | 0.181 | 30.50 ± 3.21 |
| BE (mmol/L) | −19.70 ± 4.75 | −17.82 ± 6.94 | 0.230 | 4.79 ± 3.47 |
| Na+ (mmol/L) | 134.98 ± 8.19 | 133.00 ± 7.37 | 0.389 | 140.65 ± 3.77 |
| K+ (mmol/L) | 4.80 ± 1.42a | 5.71 ± 1.85a | 0.044 | 5.45 ± 0.72 |
| Cl− (mmol/L) | 106.95 ± 8.35 | 105.88 ± 7.88 | 0.650 | 98.86 ± 2.31 |
| AG (mmol/L) | 22.67 ± 3.71 | 21.02 ± 3.39 | 0.116 | 16.74 ± 0.56 |
| SID (mmol/L) | 32.83 ± 4.64 | 32.83 ± 4.37 | 0.997 | 47.24 ± 3.61 |
| Glucose (mg/dL) | 96.89 ± 52.61 | 89.47 ± 26.29 | 0.601 | 108.95 ± 16.18 |
| BUN (mg/dL) | 47.02 ± 36.18 | 58.47 ± 34.78 | 0.267 | 8.28 ± 2.95 |
Data are presented as the mean ± SD.
Rota, diarrheic calves infected with rotavirus; Crypt, diarrheic calves infected with Cryptosporidium parvum; Healthy, clinically healthy calves without diarrhea; tCO2, total carbon dioxide; pCO2, partial pressure of carbon dioxide; HCO3 −, bicarbonate; BE, base excess; AG, anion gap; SID, strong ion difference; BUN, blood urea nitrogen.
ap < 0.05.
Stronger correlations of pH were observed with HCO3 − (r = 0.749, p < 0.001) and BE (r = 0.840, p < 0.001) in calves infected with C. parvum than in calves infected with rotavirus or clinically healthy calves. Calves infected with C. parvum also showed a stronger correlation (r = 0.788, p < 0.05) between HCO3 − and SID in both groups. The correlation of Na+ was positively strong with Cl−, but negatively moderate with K+ in both diarrheic calves. Between the two groups, the correlation of Na+ was stronger with Cl−, while weaker with K+ in calves infected with rotavirus (Table 3).
Table 3. Pearson’s correlation analysis of the acidity, and electrolyte in the blood samples from rotavirus or Cryptosporidium parvum infected calves.
| Parameter | Rota (n = 44) | Crypt (n = 17) | Healthy (n = 57) |
|---|---|---|---|
| pH – HCO3 − | 0.598c | 0.749c | 0.720c |
| pH – BE− | 0.759c | 0.840c | 0.814c |
| HCO3 − – SID | 0.626c | 0.788a | 0.245 (p = 0.066) |
| Na+ – K+ | −0.501c | −0.551a | 0.365b |
| Na+ – Cl− | 0.847c | 0.783c | 0.487c |
Rota, diarrheic calves infected with rotavirus; Crypt, diarrheic calves infected with Cryptosporidium parvum; Healthy, clinically healthy calves without diarrhea; HCO3 −, bicarbonate; BE, base excess; SID, strong ion difference.
ap < 0.05; bp < 0.01; cp < 0.001.
Serum protein profile analysis
Calves infected with C. parvum had higher levels of total protein than those infected with rotavirus due to globulin. Albumin levels were similar, but Globulin levels were significantly different. Both diarrheic groups had low albumin level (< 2.5 g/dL), and high levels of globulins (α1-, α2-, and β-globulin). However, the levels of globulin were 1.18 times and α2-globulin were 1.62 times higher in calves infected with C. parvum than those infected with rotavirus (p < 0.05). The albumin/globulin ratio was lower in calves infected with C. parvum than in those infected with rotavirus. The values of haptoglobin and serum amyloid A in both diarrheic groups were higher, but were not significantly different between the two groups (Table 4).
Table 4. Serum protein parameters of the rotavirus- or Cryptosporidium parvum- infected calves.
| Parameter | Rota (n = 40) | Crypt (n = 15) | p value | Healthy (n = 57) |
|---|---|---|---|---|
| Total protein (g/dL) | 6.02 ± 1.03 | 6.50 ± 0.94 | 0.119 | 6.24 ± 0.85 |
| Albumin (g/dL) | 2.43 ± 0.55 | 2.23 ± 0.39 | 0.207 | 3.19 ± 0.47 |
| Globulin (g/dL) | 3.60 ± 0.88a | 4.27 ± 1.05a | 0.018 | 3.06 ± 0.92 |
| Albumin/Globulin ratio | 0.71 ± 0.23 | 0.57 ± 0.27 | 0.061 | 1.15 ± 0.53 |
| α1-globulin (g/dL) | 0.93 ± 0.38 | 0.94 ± 0.39 | 0.923 | 0.80 ± 0.04 |
| α2-globulin (g/dL) | 0.95 ± 0.57a | 1.51 ± 0.86a | 0.030 | 0.37 ± 0.02 |
| β-globulin (g/dL) | 0.99 ± 0.34 | 0.96 ± 0.28 | 0.717 | 0.93 ± 0.03 |
| γ-globulin (g/dL) | 0.73 ± 0.58 | 0.86 ± 0.42 | 0.403 | 0.96 ± 0.09 |
| Haptoglobin (mg/dL) | 104.2 ± 71.0 | 130.0 ± 80.2 | 0.238 | 11.95 ± 0.59 |
| Serum amyloid A (mg/L) | 227.4 ± 89.7 | 246.5 ± 68.9 | 0.447 | 152.6 ± 95.4 |
Data are presented as the mean ± SD.
Rota, diarrheic calves infected with rotavirus; Crypt, diarrheic calves infected with Cryptosporidium parvum; Healthy, clinically healthy calves without diarrhea.
ap < 0.05.
Total protein was strongly correlated with globulin in calves (r > 0.7, p < 0.001). However, a strong correlation between globulin and α2-globulin was identified only in calves infected with C. parvum, correlation between α2-globulin and haptoglobin was strong in calves infected with C. parvum (r = 0.805, p < 0.001), but was moderate in calves infected with rotavirus (r = 0.542, p < 0.001). Meanwhile, the correlation between globulin and γ-globulin in calves infected with C. parvum (r = 0.656, p < 0.01) was moderate and lower than that in calves infected with rotavirus (r = 0.760, p < 0.001; Fig. 1).
Fig. 1. Correlations among the various types of serum proteins (Rota: 40, Crypt: 15, Healthy: 57). (A) Correlation between total protein and globulin. (B) Correlation between globulin and α2-globulin. (C) Correlation between globulin and γ-globulin. (D) Correlation between α2-globulin and haptoglobin.
Rota, diarrheic calves infected with rotavirus; Crypt, diarrheic calves infected with Cryptosporidium parvum; Healthy, clinically normal calves without diarrhea.
ap < 0.01; bp < 0.001.
DISCUSSION
This study aimed to identify the differences of acid-base, electrolyte, and biochemistry of the blood and serum proteins in diarrheic Hanwoo calves that were severely infected with rotavirus or C. parvum. Despite the similar pH, the levels and correlations of acid-base, electrolyte, and serum proteins were different. Calves infected with C. parvum showed high levels of tCO2, pCO2, HCO3 −, BE, and K+, compared to those infected with rotavirus. Calves infected with C. parvum had stronger correlations between pH and HCO3 −, and pH and BE, and Na+ and K+, but a weaker correlation between Na+ and Cl− than those infected with rotavirus. In terms of serum proteins, the level of total protein was markedly high in calves infected with C. parvum, which was associated with significantly elevated globulin and α2-globulin.
Rotavirus and C. parvum evoke diarrhea through malabsorption and secretory components [7,9,19]. Both pathogens cause intestinal villus atrophy, and stunted intestinal villi lead to malabsorption [7,19,20]. As rotavirus or C. parvum changes the villus epithelium functions, the crypt cell undergoes hyperplasia [8,9]. The mechanism by which rotavirus causes secretory diarrhea is unclear, but the enteric nervous system and viral enterotoxin play central roles [7,9]. The absorption of Na+ and glucose decreases and the secretion of Cl− increases due to the rotaviral secretory mechanism, which exacerbates the dehydration of the host [7,21]. Elevated prostaglandin production, acting on the enteric nervous system and enterocytes, mediates secretory diarrhea by C. parvum, in which the secretion of Cl− or HCO3 − increases and the absorption of NaCl decreases [7,8,19,22]. In the current study, calves infected with rotavirus showed low levels of tCO2, pCO2, HCO3 −, BE, and correlation coefficients of pH – HCO3 − and pH – BE, compared to those infected with C. parvum, despite increased HCO3 − secretion. The result is even more interesting, considering that much of the HCO3 −, remaining in the intestine lumen after the neutralization of stomach HCl, is reabsorbed in the ileum and colon [23]. The reason for this is unclear, but two mechanisms are thought to lead to lower bicarbonate levels in rotavirus infection than in C. parvum infection. One might be the difference in their preferential sites. Rotavirus infection is concentrated in the proximal small intestine, whereas C. parvum infection is most pronounced in the distal small intestine [8,21,24]. Glucose, amino acids, and lipids are mainly absorbed in the duodenum and jejunum [23,25,26,27]. Rotavirus infection reduces the activity of digestive enzymes [28,29,30]. Therefore, rotavirus may cause severe malnutrition in calves compared to C. parvum. Malnutrition increases the reaction of non-volatile acids with bicarbonate [23]. Non-volatile acids are not measured, while they lower bicarbonate level in the blood [27]. This might cause lower bicarbonate levels in calves infected with rotavirus than in those infected with C. parvum. The other mechanism might be the elevated secretion of secretin. Secretin in the duodenum and upper jejunum, stimulates bicarbonate secretion [23]. A delayed gastric emptying time, which is observed in rotavirus infection, might be induced by elevated gastrointestinal hormone secretions, including secretin [31]. Increased secretion of secretin might lead to increased bicarbonate loss.
All diarrheic calves lose potassium in the feces, which results in potassium deficiency throughout the body [32]. In general, hyperkalemia is most often observed in neonatal calves with diarrhea due to metabolic acidosis and dehydration, where increased hydrogen ion exchange with potassium ions and reduced renal elimination of potassium occurs [27,33]. However, excessive loss of potassium due to diarrhea can lead to hypokalemia and metabolic acidosis [1]. In this study, compared to clinically healthy calves, calves with rotavirus showed lower potassium levels despite metabolic acidosis, whereas those with C. parvum showed higher potassium levels. The difference might be due to the degree of potassium loss in the feces. Secretion of K+ at high rates was observed in piglet jejunum infected with rotavirus [34]. In human children, rotavirus-positive diarrhea had higher fecal K+ losses than non-rotavirus diarrhea [35]. The imbalance between the secretion and absorption of K+ in the crypt-type epithelium is speculated to lead to high K+ loss in the feces [34]. Calves might have similar mechanisms to those of children and piglets in terms of fecal K+ secretion, which contributes to hypokalemia in rotavirus infection.
Total protein levels should be interpreted with the albumin/globulin ratio. Dehydration results in hyperproteinemia with the reference interval of the albumin/globulin ratio [13,36]. In this study, diarrheic calves showed a decreased albumin/globulin ratio (low levels of albumin and high levels of globulin), compared to clinically healthy calves. Hypoalbuminemia is associated with intestinal malabsorption, malnutrition, and inflammation, and hyperglobulinemia with increased alpha globulin is associated with infection and inflammation. Albumin is a negative acute phase protein while alpha globulin is a positive acute phase protein in inflammation [13,37]. Our finding in this study corresponds to previous studies. Diarrheic calves infected with either rotavirus or C. parvum led to hypoalbuminemia and hyperglobulinemia with elevated α-globulin levels. In addition, acute-phase proteins such as haptoglobin and serum amyloid A were synthesized following infection with either of the pathogens. Interestingly, calves infected by C. parvum had higher α2-globulin levels, including haptoglobin than those infected with rotavirus. The magnitude and type of immune responses depend on the pathogen, and the type of globulins produced also differs according to the type of pathogen [38]. Acute phase responses are more pronounced in bacterial infections than in viral infections [39]. More production of α2-globulin including haptoglobin may be an immunological characteristic of C. parvum infection.
The current study has a few limitations. Mild cases of rotavirus and C. parvum infection were not included. More cases should have been investigated to compare single infection with concurrent infections of rotavirus and C. parvum. In addition, diarrheic calves by the degree of clinical symptoms (dehydration or duration of manifestation) were not subdivided. Further studies are required to elucidate the impact of these two pathogens in details. However, to the best of our knowledge, the current study is the first field study that compares acid-base, electrolyte, and biochemical parameters in calves with severe diarrhea caused by rotavirus or C. parvum infections. Even though rotavirus and C. parvum cause malabsorptive and secretory diarrhea in similar-aged calves, blood parameters were different.
In conclusion, the rotavirus-infected calves significantly low levels of potassium, globulin, and α2-globulin and weak correlations between pH and HCO3 −, pH and base excess, HCO3 − and SID, globulin and α2-globulin, α2-globulin and haptoglobin, compared to C. parvum-infected calves. Our findings would contribute to understandings of the host responses to rotavirus and C. parvum infections in the field, differential diagnosis, and the establishment of well-informed treatment strategies for both pathogens in diarrheic neonatal calves.
ACKNOWLEDGEMENTS
We would like to thank the farm owners and caretakers for helping with the collection of samples.
Footnotes
Funding: This research was supported by a National Research Foundation of Korea (NRF) grant funded by the Korean government (MSIT) (No. NRF2021R1A2C100517111).
Conflict of Interest: The authors declare no conflicts of interest.
- Conceptualization: Ha S, Park J.
- Data curation: Ha S, Park J.
- Formal analysis: Ha S, Kang S.
- Funding acquisition: Park J.
- Investigation: Park KM, Ku JY, Choi KS.
- Methodology: Ha S, Kang S.
- Project administration: Park J.
- Resources: Park J.
- Software: Ha S.
- Supervision: Park J.
- Validation: Ha S.
- Visualization: Ha S.
- Writing - original draft: Ha S.
- Writing - review & editing: Park J.
References
- 1.Lim S. Approach to hypokalemia. Acta Med Indones. 2007;39(1):56–64. [PubMed] [Google Scholar]
- 2.Todd CG, Millman ST, McKnight DR, Duffield TF, Leslie KE. Nonsteroidal anti-inflammatory drug therapy for neonatal calf diarrhea complex: effects on calf performance. J Anim Sci. 2010;88(6):2019–2028. doi: 10.2527/jas.2009-2340. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 3.Cho YI, Yoon KJ. An overview of calf diarrhea - infectious etiology, diagnosis, and intervention. J Vet Sci. 2014;15(1):1–17. doi: 10.4142/jvs.2014.15.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 4.Meganck V, Hoflack G, Piepers S, Opsomer G. Evaluation of a protocol to reduce the incidence of neonatal calf diarrhoea on dairy herds. Prev Vet Med. 2015;118(1):64–70. doi: 10.1016/j.prevetmed.2014.11.007. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 5.Saif L, Rosen B, Parwani A. Diagnostic Veterinary Virology: A Practitioner’s Guide. Baltimore: Williams and Wilkins; 1991. Bovine rotavirus; pp. 126–130. [Google Scholar]
- 6.Silverlås C, Bosaeus-Reineck H, Näslund K, Björkman C. Is there a need for improved Cryptosporidium diagnostics in Swedish calves? Int J Parasitol. 2013;43(2):155–161. doi: 10.1016/j.ijpara.2012.10.009. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 7.Foster DM, Smith GW. Pathophysiology of diarrhea in calves. Vet Clin North Am Food Anim Pract. 2009;25(1):13–36. doi: 10.1016/j.cvfa.2008.10.013. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 8.Laurent F, McCole D, Eckmann L, Kagnoff MF. Pathogenesis of Cryptosporidium parvum infection. Microbes Infect. 1999;1(2):141–148. doi: 10.1016/s1286-4579(99)80005-7. [DOI] [PubMed] [Google Scholar]
- 9.Ramig RF. Pathogenesis of intestinal and systemic rotavirus infection. J Virol. 2004;78(19):10213–10220. doi: 10.1128/JVI.78.19.10213-10220.2004. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 10.Enemark HL, Bille-Hansen V, Lind P, Heegaard PM, Vigre H, Ahrens P, et al. Pathogenicity of Cryptosporidium parvum--evaluation of an animal infection model. Vet Parasitol. 2003;113(1):35–57. doi: 10.1016/S0304-4017(03)00034-7. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 11.Estes MK, Atmar RL. In: Microbial Pathogenesis and the Intestinal Epithelial Cell. Hecht GA, editor. Hoboken: ASM Press; 2003. Viral pathogens of the intestine; pp. 525–545. [Google Scholar]
- 12.Lee SH, Choi EW, Kim D. Relationship between the values of blood parameters and physical status in Korean native calves with diarrhea. J Vet Sci. 2020;21(2):e17. doi: 10.4142/jvs.2020.21.e17. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 13.Latimer KS. Duncan and Prasse’s Veterinary Laboratory Medicine: Clinical Pathology. Hoboken: John Wiley & Sons; 2011. [Google Scholar]
- 14.Tîrziu E. Acute-phase proteins in immune response. Lucrări Ştiinłifice Medicină Veterinară. 2009;42(1):329–339. [Google Scholar]
- 15.Choi KS, Kang JH, Cho HC, Yu DH, Park J. Changes in serum protein electrophoresis profiles and acute phase proteins in calves with diarrhea. Can J Vet Res. 2021;85(1):45–50. [PMC free article] [PubMed] [Google Scholar]
- 16.Cho YI, Kim WI, Liu S, Kinyon JM, Yoon KJ. Development of a panel of multiplex real-time polymerase chain reaction assays for simultaneous detection of major agents causing calf diarrhea in feces. J Vet Diagn Invest. 2010;22(4):509–517. doi: 10.1177/104063871002200403. [DOI] [PubMed] [Google Scholar]
- 17.Mahlum CE, Haugerud S, Shivers JL, Rossow KD, Goyal SM, Collins JE, et al. Detection of bovine viral diarrhea virus by TaqMan reverse transcription polymerase chain reaction. J Vet Diagn Invest. 2002;14(2):120–125. doi: 10.1177/104063870201400205. [DOI] [PubMed] [Google Scholar]
- 18.Sayers RG, Kennedy A, Krump L, Sayers GP, Kennedy E. An observational study using blood gas analysis to assess neonatal calf diarrhea and subsequent recovery with a European Commission-compliant oral electrolyte solution. J Dairy Sci. 2016;99(6):4647–4655. doi: 10.3168/jds.2015-10600. [DOI] [PubMed] [Google Scholar]
- 19.Wyatt CR, Riggs MW, Fayer R. Cryptosporidiosis in neonatal calves. Vet Clin North Am Food Anim Pract. 2010;26(1):89–103. doi: 10.1016/j.cvfa.2009.10.001. [DOI] [PubMed] [Google Scholar]
- 20.Heller MC, Chigerwe M. Diagnosis and treatment of infectious enteritis in neonatal and juvenile ruminants. Vet Clin North Am Food Anim Pract. 2018;34(1):101–117. doi: 10.1016/j.cvfa.2017.08.001. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 21.Lundgren O, Svensson L. Pathogenesis of rotavirus diarrhea. Microbes Infect. 2001;3(13):1145–1156. doi: 10.1016/S1286-4579(01)01475-7. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 22.Gookin JL, Nordone SK, Argenzio RA. Host responses to Cryptosporidium infection. J Vet Intern Med. 2002;16(1):12–21. doi: 10.1892/0891-6640(2002)016<0012:hrtci>2.3.co;2. [DOI] [PubMed] [Google Scholar]
- 23.Cunningham JG, Klein BG. Cunningham’s Textbook of Veterinary Physiology. 5th ed. St. Louis: Elsevier Health Sciences; 2013. [Google Scholar]
- 24.Hodges R. Cryptosporidiosis in calves. J Clin Immunol. 2002;124(6):1152–1160. [Google Scholar]
- 25.Harmon DL. Understanding starch utilization in the small intestine of cattle. Asian-Australas J Anim Sci. 2009;22(7):915–922. [Google Scholar]
- 26.Shirazi-Beechey S, Wood IS, Dyer J, Scott D, King TP. In: Ruminant Physiology: Digestion, Metabolism, Growth and Reproduction. Engelhardt WV, Leonhard S, Breves G, Giesecke D, editors. Stuttgart: Ferdinand Enke Verlag; 1995. Intestinal sugar transport in ruminants; pp. 117–133. [Google Scholar]
- 27.Reece WO, Erickson HH, Goff JP, Uemura EE. Dukes’ Physiology of Domestic Animals. 13th ed. Ames: John Wiley & Sons; 2015. [Google Scholar]
- 28.Davidson GP, Gall DG, Petric M, Butler DG, Hamilton JR. Human rotavirus enteritis induced in conventional piglets. Intestinal structure and transport. J Clin Invest. 1977;60(6):1402–1409. doi: 10.1172/JCI108901. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 29.Collins J, Starkey WG, Wallis TS, Clarke GJ, Worton KJ, Spencer AJ, et al. Intestinal enzyme profiles in normal and rotavirus-infected mice. J Pediatr Gastroenterol Nutr. 1988;7(2):264–272. doi: 10.1097/00005176-198803000-00017. [DOI] [PubMed] [Google Scholar]
- 30.Brunet JP, Cotte-Laffitte J, Linxe C, Quero AM, Géniteau-Legendre M, Servin A. Rotavirus infection induces an increase in intracellular calcium concentration in human intestinal epithelial cells: role in microvillar actin alteration. J Virol. 2000;74(5):2323–2332. doi: 10.1128/jvi.74.5.2323-2332.2000. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 31.Crawford SE, Ramani S, Tate JE, Parashar UD, Svensson L, Hagbom M, et al. Rotavirus infection. Nat Rev Dis Primers. 2017;3(1):17083. doi: 10.1038/nrdp.2017.83. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 32.Lewis LD, Phillips RW. Water and electrolyte losses in neonatal calves with acute diarrhea. A complete balance study. Cornell Vet. 1972;62(4):596–607. [PubMed] [Google Scholar]
- 33.Sweeney RW. Treatment of potassium balance disorders. Vet Clin North Am Food Anim Pract. 1999;15(3):609–617. doi: 10.1016/s0749-0720(15)30166-3. [DOI] [PubMed] [Google Scholar]
- 34.Woodard JP, Chen W, Keku EO, Liu SC, Lecce JG, Rhoads JM. Altered jejunal potassium (Rb+) transport in piglet rotavirus enteritis. Am J Physiol. 1993;265(2 Pt 1):G388–G393. doi: 10.1152/ajpgi.1993.265.2.G388. [DOI] [PubMed] [Google Scholar]
- 35.Rerksuppaphol S, Rerksuppaphol L. Prevalence and clinical manifestations of rotavirus diarrhea in children of rural area of Thailand. Int J Collab Res Intern Med Public Health. 2011;3(9):695–702. [Google Scholar]
- 36.Barger AM, MacNeill AL. Clinical Pathology and Laboratory Techniques for Veterinary Technicians. John Wiley & Sons; 2015. [Google Scholar]
- 37.Thrall MA, Weiser G, Allison RW, Campbell TW. Veterinary Hematology and Clinical Chemistry. Ames: John Wiley & Sons; 2012. [Google Scholar]
- 38.Tóthová C, Mihajlovičová X, Nagy O. In: Ruminants - The Husbandry, Economic and Health Aspects. Abubakar M, Iqbal A, Kabir A, Manzoor S, editors. London: IntechOpen Ltd.; 2018. The use of serum proteins in the laboratory diagnosis of health disorders in ruminants; pp. 105–146. [Google Scholar]
- 39.Ulutas B, Tan T, Ulutas PA, Bayramli G. Haptoglobin and serum amyloid A responses in cattle persistently infected with bovine viral diarrhea virus. Acta Sci Vet. 2011;39(3):973. [Google Scholar]