Upregulation of PD-L1 Expression by Prostaglandin E2 and the Enhancement of IFN-γ by Anti-PD-L1 Antibody Combined With a COX-2 Inhibitor in Mycoplasma bovis Infection.

Bovine mycoplasmosis caused by Mycoplasma bovis results in pneumonia and mastitis in cattle. We previously demonstrated that the programmed death 1 (PD-1)/PD-ligand 1 (PD-L1) pathway is involved in immune dysfunction during M. bovis infection and that prostaglandin E2 (PGE2) suppressed immune responses and upregulated PD-L1 expression in Johne's disease, a bacterial infection in cattle. In this study, we investigated the role of PGE2 in immune dysfunction and the relationship between PGE2 and the PD-1/PD-L1 pathway in M. bovis infection. In vitro stimulation with M. bovis upregulated the expressions of PGE2 and PD-L1 presumably via Toll-like receptor 2 in bovine peripheral blood mononuclear cells (PBMCs). PGE2 levels of peripheral blood in infected cattle were significantly increased compared with those in uninfected cattle. Remarkably, plasma PGE2 levels were positively correlated with the proportions of PD-L1+ monocytes in M. bovis-infected cattle. Additionally, plasma PGE2 production in infected cattle was negatively correlated with M. bovis-specific interferon (IFN)-γ production from PBMCs. These results suggest that PGE2 could be one of the inducers of PD-L1 expression and could be involved in immunosuppression during M. bovis infection. In vitro blockade assays using anti-bovine PD-L1 antibody and a cyclooxygenase 2 inhibitor significantly upregulated the M. bovis-specific IFN-γ response. Our study findings might contribute to the development of novel therapeutic strategies for bovine mycoplasmosis that target PGE2 and the PD-1/PD-L1 pathway.

Immunosuppression in Cows following Intramammary Infusion of Mycoplasma bovis.

Mycoplasma bovis is a destructive pathogen that causes large economic losses in rearing cattle for beef and dairy worldwide. M. bovis causes suppression of and evades the host immune response; however, the mechanisms of host immune function involved in M. bovis mastitis have not been elucidated. The purpose of this study was to elucidate the characteristics of the bovine immune response to mycoplasmal mastitis. We evaluated the responsiveness of the bovine mammary gland following infusion of M. bovis Somatic cell counts and bacterial counts in milk from the infected quarter were increased. However, the proliferation of peripheral blood mononuclear cells (blood MNCs) and mononuclear cells isolated from M. bovis-stimulated mammary lymph nodes (lymph node MNCs) did not differ from that in the unstimulated cells. Transcriptome analysis revealed that the mRNA levels of innate immune system-related genes in blood MNCs, complement factor D (CFD), ficolin 1 (FCN1), and tumor necrosis factor superfamily member 13 (TNFSF13) decreased following intramammary infusion of M. bovis The mRNA levels of immune exhaustion-related genes, programmed cell death 1 (PD-1), programmed cell death-ligand 1 (PD-L1), lymphocyte activation gene 3 (LAG3), and cytotoxic T-lymphocyte-associated protein 4 (CTLA4) of milk mononuclear cells (milk MNCs) in the infected quarter were increased compared with those before infusion. Increase in immune exhaustion-related gene expression and decrease in innate immune response-related genes of MNCs in quarters from cows were newly characterized by M. bovis-induced mastitis. These results suggested that M. bovis-induced mastitis affected the immune function of bovine MNCs, which is associated with prolonged duration of infection with M. bovis.

Bovine milk transcriptome analysis reveals microRNAs and RNU2 involved in mastitis.

Mastitis is a common inflammatory infectious disease in dairy cows. To understand the microRNA (miRNA) expression profile changes during bovine mastitis, we undertook a genome-wide miRNA study of normal milk and milk that tested positive on the California mastitis test for bovine mastitis (CMT+). Twenty-five miRNAs were differentially expressed (23 miRNAs upregulated and two downregulated) during bovine mastitis relative to their expression in normal milk. Upregulated mature miR-1246 probably derived from a U2 small nuclear RNA rather than an miR-1246 precursor. The significantly upregulated miRNA precursors and RNU2 were significantly enriched on bovine chromosome 19, which is homologous to human chromosome 17. A gene ontology analysis of the putative mRNA targets of the significantly upregulated miRNAs showed that these miRNAs were involved in binding target mRNA transcripts and regulating target gene expression, and a Kyoto Encyclopedia of Genes and Genomes pathway analysis showed that the upregulated miRNAs were predominantly related to cancer and immune system pathways. Three novel miRNAs were associated with bovine mastitis and were relatively highly expressed in milk. We confirmed that one of the novel mastitis-related miRNAs was significantly upregulated using a digital PCR system. The differentially expressed miRNAs were involved in human cancers, infections, and immune-related diseases. The genome-wide analysis of miRNA profiles in this study provides insight into bovine mastitis and inflammatory diseases. DATABASES: The miRNAseq generated for this study can be found in the Sequence Read Archive (SRA) under BioProject Number PRJNA421075 and SRA Study Number SRP126134 (https://www.ncbi.nlm.nih.gov/bioproject/PRJNA421075).

Cytokine levels of peripheral blood mononuclear cells in the clinical cases of Holstein calves infected with Mycoplasma bovis.

The immune related factors of peripheral blood mononuclear cells (PBMC) were analyzed in the clinical cases with Mycoplasma (M.) bovis infection. Seventy-eight Holstein calves in one farm were used. These calves were divided into three groups; the calves with M. bovis infection of poor outcome after treatment (Non-Recovery Group), the calves with M. bovis infection recovered (Recovery Group) and clinically healthy calves (Control Group). Blood samples were collected at days of the first medical treatment and the final treatment or euthanasia. IL-17A levels in the Non-Recovery Group were higher than those in the Recovery Group on both days. Our result suggested that the IL-17A of PBMC is an important factor to affect outcome of the calves with M. bovis infection.

Effect of Mycoplasma bovis on expression of inflammatory cytokines and matrix metalloproteinases mRNA in bovine synovial cells.

Mycoplasma bovis causes chronic arthritis in calves. Mycoplasma arthritis shows severe inflammatory reactions in joints that is commonly treated with antibiotics and results in significant economic losses in the calf industry. A previous study showed that inflammatory cytokines and matrix metalloproteinases (MMPs) produced by synovial cells promote progression of the pathophysiology of bacterial arthritis. However, the mechanism underlying the pathogenesis of bovine Mycoplasma arthritis has not been fully clarified. In this study, we examined the immunologic response of bovine synovial tissue to M. bovis. We observed significant increases in expression of interleukin (IL)-1ß, IL-6, IL-8, MMP-1, and MMP-3 mRNA in synovial tissue from Mycoplasma arthritis calves compared with tissues from normal calves. Expression of IL-6, IL-8, and MMP-1 mRNA was also induced in cultured synovial cells stimulated with M. bovis, but not expression of IL-1ß and MMP-3 mRNA. In contrast, the culture supernatant of peripheral blood mononuclear cells stimulated with M. bovis induced marked increases in the expression of IL-1ß, IL-6, IL-8, MMP-1, and MMP-3 mRNA in synovial cells. Our results indicate that inflammatory cytokines and MMPs produced by synovial cells play a key role in the pathogenesis of Mycoplasma arthritis. We suggest that interactions between synovial cells and mononuclear cells in the presence of M. bovis induce expression of these cytokines and MMPs in synovial cells, resulting in severe inflammatory reactions in the joints.

Low-field magnetic resonance imaging and computed tomography of a calf with aqueductal stenosis caused by web: comparison with normal calves.

A 6-day-old female Holstein displayed a dome-shaped skull and cardiac murmur on physical examination. Neurological abnormalities included progressive ataxia, decreased pupillary light reflex, and blindness soon after birth. On diagnostic imaging, CT identified expanded ventricles and thyroid hypoplasia on the left side. MRI detected expanded ventricles, especially in the rostral cerebrum at the mesencephalic aqueduct, compared with normal calves, so we suspected hydrocephalus causing stenosis of the mesencephalic aqueduct. Postmortem examination revealed a structure in the mesencephalic aqueduct resembling the "web" type of aqueductal stenosis described in humans. This case report indicates the utility of describing mesencephalic aqueductal stenosis by web and detection of other malformations on CT and MRI for antemortem diagnosis in calves.

Bacteriological survey of bulk tank milk from dairy farms in Montero, Santa Cruz, Bolivia.

BACKGROUND AND AIM: Recently, bacterial surveys for mastitis-causing pathogens in bulk tank milk (BTM) have been conducted in several countries worldwide. However, no such surveys have been reported from Bolivia. Therefore, the present study aimed to estimate the prevalence of mastitis pathogens in BTM from dairy farms in Montero, Santa Cruz, Bolivia. MATERIALS AND METHODS: Between July 2016 and August 2017, a total of 43 BTM samples were collected from 3264 cows to determine bulk tank somatic cell counts (BTSCC) and identify mastitis-causing bacteria. BTSCC was classified as follows: <100×103, 100-500×103, 500-1000×103, and >1000×103 cells/mL. RESULTS: Mastitis-causing pathogens identified by agar medium cultures included Bacillus spp., coagulase-negative staphylococci (CNS), coliforms, Staphylococcus aureus (SA), streptococci, and other species. The proportions of BTSCC of <100×103, 200-500×103, 500-1000×103, and >1000×103 cells/ml were 0%, 37%, 51%, and 12%, respectively. The proportions of coliforms, streptococci, CNS, Bacillus spp., SA, and others detected in BTM were 33%, 30%, 16%, 7%, 2%, and 16%, respectively. CONCLUSION: Although the herd prevalence of contagious mastitis-causing pathogens, such as SA, in BTM was low, increased BTSCC were identified in Montero, Santa Cruz, Bolivia.

Inflammation-related microRNA expression level in the bovine milk is affected by mastitis.

MicroRNA (miRNA) in tissue and liquid samples have been shown to be associated with many diseases including inflammation. We aimed to identify inflammation-related miRNA expression level in the bovine mastitis milk. Expression level of inflammation-related miRNA in milk from mastitis-affected and normal cows was analyzed using qPCR. We found that expression level of miR-21, miR-146a, miR-155, miR-222, and miR-383 was significantly upregulated in California mastitis test positive (CMT+) milk. We further analyzed these miRNA using a chip-based QuantStudio Digital PCR System. The digital PCR results correlated with those of qPCR, demonstrating upregulation of miR-21, miR-146a, miR-155, miR-222, and miR-383 in CMT+ milk. In conclusion, we identified miRNA that are upregulated in CMT+ milk. These miRNA exhibited sensitivity and specificity greater than 80% for differentiating between CMT+ milk and normal milk. Our findings suggest that inflammation-related miRNA expression level in the bovine milk was affected by mastitis, and miRNA in milk have potential for use as biomarkers of bovine mastitis.

Increase of cells expressing PD-1 and PD-L1 and enhancement of IFN-γ production via PD-1/PD-L1 blockade in bovine mycoplasmosis.

INTRODUCTION: Bovine mycoplasma, chiefly Mycoplasma bovis, is a pathogen that causes pneumonia, mastitis, arthritis, and otitis media in cattle. This pathogen exerts immunosuppressive effects, such as the inhibition of interferon production. However, the mechanisms involved in bovine mycoplasmosis have not been fully elucidated. In this study, we investigated the role of the programmed death-1 (PD-1)/programmed death-ligand 1 (PD-L1) pathway in immunosuppression in bovine mycoplasmosis. METHODS: In the initial experiments, we used enzyme-linked immunosorbent assay to measure interferon-γ (IFN-γ) from peripheral blood mononuclear cells (PBMCs) isolated from cattle with mycoplasmosis. RESULTS: Expectedly, IFN-γ production significantly decreased in cattle with mycoplasmosis compared with that in clinically healthy cattle. Concomitantly, flow cytometric analysis revealed that the proportions of PD-1+ CD4+ and PD-L1+ CD14+ cells significantly increased in peripheral blood of the infected cattle. Interestingly, the number of PD-1+ CD4+ and PD-1+ CD8+ T cells were negatively correlated with IFN-γ production from PBMCs in bovine mycoplasmosis. Additionally, blockade of the PD-1/PD-L1 pathway in vitro by anti-bovine PD-1- and anti-bovine PD-L1 antibodies significantly upregulated the production of IFN-γ from anti-mycoplasma-specific cells. CONCLUSIONS: These results suggest that the PD-1/PD-L1 pathway could be involved in immune exhaustion of bovine mycoplasma-specific T cells. In conclusion, our study opens up a new perspective in the therapeutic strategy for bovine mycoplasmosis by targeting the immunoinhibitory receptor pathways.

Particle-Induced X-ray Emission Analysis of Zierum Trace and Major Elements in Cattle with Acute Coliform Mastitis.

The aim of the present study was to examine the applicability of the direct determination of trace and major element concentrations in serum samples collected from Holstein dairy cattle with acute coliform mastitis (n = 53) compared with a healthy control group (n = 39). Twenty-eight elements (Na, Mg, Al, Si, S, Cl, K, Ca, Ti, V, Cr, Mn, Fe, Ce, Ni, Cu, Zn, Ga, As, Se, Br, Rb, Sr, Y, Zr, Nb, Mo, and Pb) were detected by particle-induced X-ray emission (PIXE). Significant differences were observed in serum K, Fe, Zn, and Br concentrations, but not in those of the remaining twenty-four elements. Furthermore, serum Fe concentrations (0.751 ± 0.583 µg/ml, n = 18) were significantly lower in dairy cattle with a poor prognosis than in those with a good prognosis (0.945 ± 0.393 µg/ml, n = 35, P < 0.05) and healthy controls (1.458 ± 0.391 µg/ml, n = 39, P < 0.01). We proposed a diagnostic cut-off point for serum Fe concentrations of <0.82 µg/ml based on receiver operating characteristic (ROC) curves in order to identify cattle with a poor prognosis. The results of the present study indicated that assessing the elemental composition of serum, particularly iron, is a promising prognostic tool for determining the outcomes of cattle with severe acute coliform mastitis.

Sero-epidemiological analysis of vertical transmission relative risk of Borna disease virus infection in dairy herds.

Borna disease virus (BDV) is a virus that causes a neurological disease in domestic animals, including a variety of animal species in Japan. Few studies have examined the mode of transmission of this virus in cattle, and the exact mechanisms underlying the transmission of the virus need to be elucidated. This study aimed to examine the contribution of vertical transmission of the virus, which occurs when the virus is transmitted from the mother to offspring during gestation or birth. We used an epidemiological approach. The relative risk (RR) was calculated for cattle born to BDV sero-positive cows from farms with a higher within-herd prevalence of BDV (56.8%). We tested the sera of 1,122 dairy cattle from 24 dairy herds in Hokkaido Prefecture, Japan, for BDV infection using the ELISA and western blotting method. The overall level of BDV sero-prevalence was 22.1%. Seroprevalence was significantly higher in closed-breeding herds that do not have buying in cows (39.7%) than in farms that restock cattle by buying in cows (4.4%, P<0.01). The overall RR of BDV vertical transmission from infected mothers to their daughters was 1.86 (95% confidence interval (CI): 1.54-2.56). Our results show that vertical transmission contributes significantly to BDV transmission in the farms tested in this study.

Primary ischial osteosarcoma occupying the pelvic cavity in a Japanese Black cow.

A 10-year-old Japanese Black cow presented with a swelling of the right femur, and a hard, large mass occupied the pelvic cavity. The mass strongly adhered to the visceral surface of the ischium and had posteriorly invaded among the right femoral muscles. Histologically, the mass was composed of neoplastic osteoblasts and exhibited osteoid and immature trabecular bone production. In the region where the mass adhered to the ischium, neoplastic cells were continuously proliferating into the medullary cavity. Tumor emboli were observed in the small vessels of the femoral muscles and lungs. Based on these findings, the mass was diagnosed as an osteosarcoma and considered to have arisen from the ischium.

Testicular yolk sac tumor of myxomatous, reticular, and polyvesicular vitelline type in a newborn calf.

Yolk sac tumors (YSTs) are rare neoplasms of germ cell origin. In humans, the tumors primarily occur in the testes or ovaries, but occasionally develop at other sites. The neoplastic cells of YSTs form many histological patterns resembling embryonal structures, and the World Health Organization classification lists 11 such patterns: reticular, macrocystic, endodermal sinus, papillary, solid, glandular-alveolar, myxomatous, sarcomatoid, polyvesicular vitelline, hepatoid, and parietal. Among domestic animals, only 2 cases of YST, which were of testicular and abdominal cavity origin, have been reported in calves. In both cases, neoplastic cells had epithelial properties and disseminated metastases in the abdomen. In the present study, the enlarged testis of a newborn calf, which was subsequently diagnosed as YST and exhibited myxomatous, reticular, and polyvesicular vitelline histological patterns, is described. There was no metastasis in this case, and histological and immunohistochemical features varied from previous cases of YST.

Changes in leukocyte populations of cows with milk fever or displaced abomasum after calving.

Most of the metabolic diseases of dairy cows occur within the first 2 wk after calving, and cows with a metabolic disease are prone to infectious diseases. Although metabolic diseases are generally recognized as a risk factor for infectious diseases owing to the associated decrease in immune function, the difference in immune status between cows with milk fever (MF) or displaced abomasum (DA) during the lactation period has not been clarified. Therefore, the peripheral blood leukocyte populations in 38 multiparous Holstein cows from 1 herd were analyzed after calving. The cows were divided into 3 groups according to health: 21 cows that remained clinically healthy throughout the experimental period (control group), 9 cows that had MF on the day of calving, and 8 cows with an onset of DA within 4 wk after calving. The T- and B-cell numbers were lowest at week 0, and they increased gradually after calving. There was no significant difference between the 3 groups in the number of each subset of leukocytes on the day of calving, but the number of CD8⁺ T-cells was significantly lower in the MF and DA groups than in the control group at week 1. The numbers of CD4⁺, CD8⁺, and WC1⁺ T-cells tended to be lower in the DA group than in control group from weeks 4 to 12, a tendency not observed in the MF group. These data suggest that when cows have DA around the time of calving, their lymphocyte numbers remain lower until 12 wk after calving.

La plupart des maladies métaboliques des vaches laitières se produisent à l'intérieur des 2 premières semaines après le vêlage, et les vaches avec une maladie métabolique sont sujettes aux maladies infectieuses. Bien que les maladies métaboliques soient généralement reconnues comme un facteur de risque pour les maladies infectieuses à cause de la diminution de la fonction immunitaire qui y est associée, la différence dans le statut immunitaire entre des vaches avec une fièvre du lait (MF) ou un déplacement de caillette (DA) durant la période de lactation n'a pas été clarifiée. Ainsi, les populations des leucocytes du sang périphérique de 38 vaches Holstein multipares provenant de un troupeau ont été analysées après le vêlage. Les vaches ont été divisées en trois groupes selon leur état de santé : 21 vaches qui sont demeurées cliniquement saines durant toute la durée de la période expérimentale (groupe témoin), 9 vaches qui ont eu une MF le jour du vêlage, et 8 vaches avec un début de DA dans les 4 sem après le vêlage. Le nombre de cellules T et B étaient à son plus bas la semaine 0, et il a augmenté graduellement après le vêlage. Il n'y avait pas de différence significative entre les trois groupes dans le nombre de chaque sous-groupe de leucocytes le jour du vêlage, mais le nombre de cellules T CD8+ était significativement plus faible dans les groupes MF et DA que dans le groupe témoin à la semaine 1. Le nombre de cellules T CD4+, CD8+ et WC1+ avaient tendance à être plus faible dans le groupe DA que dans le groupe témoin à partir de la semaine 4 jusqu'à la semaine 12, une tendance non observée dans le groupe MF. Ces données suggèrent que lorsque les vaches ont un DA durant la période du vêlage, leurs dénombrements lymphocytaires demeurent faibles jusqu'à 12 sem après la mise-bas.(Traduit par Docteur Serge Messier).

Molecular-based identification of yeasts isolated from bovine clinical mastitis in Japan.

This study analyzed molecular-based identification of yeasts that associated with bovine clinical mastitis in Japan. Over 3,200 quarter milk samples from Holstein dairy cows collected in 2011 on Hokkaido and Honshu islands were examined. Yeast isolates were characterized by polymerase chain reaction amplification and sequencing of the D1/D2 region of the 26S rDNA. Molecular characterization confirmed that Candida spp. and Pichia spp. were most frequently isolated species. Our molecular analysis of mastitic milk samples demonstrated the prevalence of Pichia kudriavzevii(22/58) and Candida tropicalis(14/58). In addition, we demonstrated that molecular analysis of the D1/D2 region of the 26S rDNA is a rapid and reliable method for identifying clinically significant yeasts in dairy hygiene, including potentially new or emerging pathogenic species.

The influence of Borna disease viral infection on dairy cow reproduction.

We investigated the influence of Borna disease virus (BDV) infection on the clinical state of dairy cows. Sera from 149 cows were examined using enzyme-linked immunosorbent assay and western blotting detect antibodies to the BDV-nucleoprotein antigen. Among 149 investigated cows, 25 (16.8%) showed a positive reaction to BDV antigen. No significant difference existed in milk production or medical history between seropositive and seronegative cows. Although the estrus cycle appeared normal even in the seropositive cows, the frequency of artificial insemination and calving-to-conception intervals significantly increased in seropositive cows. Therefore, fertilization failure was recognized in the BDV-antibody positive cows.

Analysis of trace and major elements in bronchoalveolar lavage fluid of Mycoplasma bronchopneumonia in calves.

The aim of this study was to evaluate the reliability and effectiveness of direct determination of trace and major element concentrations in bronchoalveolar lavage fluid samples from Holstein calves with Mycoplasma bronchopneumonia (n = 21) and healthy controls (n = 20). The samples were obtained during bronchoscopy using a standard examination method. A total of 18 elements (aluminum, bromine, calcium, chlorine, chromium, copper, iron, potassium, magnesium, manganese, molybdenum, nickel, phosphorous, sulfur, silicon, strontium, titanium, and zinc) were detected by particle-induced X-ray emission. The average bromine, iron, potassium, magnesium, and phosphorous concentrations were higher in calves with bronchopneumonia than in controls (p < 0.05). They were found to have higher amounts of calcium and zinc, and a higher zinc-copper ratio than that in healthy calves (p < 0.001). Based on the receiver operating characteristics curves, we propose a diagnostic cutoff point for zinc-copper ratio for identification of Mycoplasma pneumonia of 8.676. Our results indicate that assessment of the elemental composition of broncholaveolar lavage fluid is a promising diagnostic tool for Mycoplasma bronchopneumonia.

Genetic analysis and development of species-specific PCR assays based on ITS-1 region of rRNA in bovine Eimeria parasites.

At present, morphological characteristics of oocyst is the only achievable method for the identification of bovine coccidia to the species level. In this study, the internal transcribed spacer 1 (ITS-1) region of ribosomal RNA genes of six bovine Eimeria species; E. alabamensis, E. auburnensis, E. bovis, E. cylindrica, E. ellipsoidalis and E. zuernii, were sequenced and analyzed the phylogenetic relationship among them. In pair-wise alignment, the sequences among the same species had high homology of over 90%. E. bovis and E. zuernii were closely related within the same cluster. This cluster and E. alabamensis were distant from major cluster of bovine coccidia that included E. auburnensis, E. cylindrica and E. ellipsoidalis. Species-specific PCR assays based on the amplification of the ITS-1 region were also developed to identify the 6 pathogens. The ITS-1 region of each Eimeria species had sufficient inter-specific sequence variation enough to design the primer sets that differentially amplified each target species. This PCR assay for the detection and differentiation of Eimeria parasite showed higher sensitivity when compared to the conventional oocyst-morphological examination. This is the first attempt for the identification of 6 bovine Eimeria parasites in the genomic level and may provide as useful methods for diagnosis and epidemiology of bovine coccidial infection.

Non-cytopathic bovine viral diarrhea virus infection inhibits differentiation of bovine neural stem/progenitor cells into astrocytes in vitro.

Bovine viral diarrhea virus (BVDV) causes fetal brain malformations in ruminants when the fetuses are infected transplacentally in mid-pregnancy. In both cytopathic and non-cytopatic virus infections, viral lytic infection in actively replicating cells and interruption of vascular integrity have been suggested as the pathogenesis, but functional disturbance of infected neural developing cells has been unclear. In this study, we examined the effect of infection with non-cytopathic BVDV2 on the differentiation of neural stem/precursor cells isolated from the bovine fetus. In the process of differentiation to three types of neural cells, neurons, astrocytes and oligodendrocytes, virus infection significantly and selectively inhibited the differentiation of neural stem/precursor cells into the astrocytic lineage. This inhibition is possibly important for the pathogenesis of congenital brain malformations associated with non-cytopathic BVDV infection.

Comparison of the alkalizing effects of bicarbonate precursors in calves with experimentally induced metabolic acidosis.

The aims of this study were to confirm whether commercial acetated Ringer's solution, which contains 28 mM of sodium acetate, is superior to commercial lactated Ringer's solution in alkalizing effects in calves with experimentally induced metabolic acidosis. Twenty calves with experimentally induced mild acidosis were intravenously administered isotonic saline, DL-lactated, L-lactated or acetated Ringer's solution at a dose of 80 ml/kg body weight (BW). The acetated Ringer's solution induced a significantly greater increase in venous HCO(3)(-) and base excess concentrations than the other fluids during the early phases of extracellular fluid replacement in mild metabolic acidosis. Therefore, the alkalizing effect of commercial acetated Ringer's solution is superior to commercial DL- and L-lactated Ringer's solution in treatment of mild metabolic acidosis in calves.