THE EFFECTIVENESS OF ZINC SUPPLEMENTED ORAL REHYDRATION SOLUTION ON FECAL OUTPUT, RECOVERY AND MORTALITY RATES IN NEONATAL HOLSTEIN CALVES WITH DIARRHEA

• Sponsoring Institution: National Institute of Food and Agriculture
• Project Status: COMPLETE
• Funding Source: ANIMAL HEALTH
• Reporting Frequency: Annual
• Accession No.: 0225976
• Project Start Date: Oct 1, 2011
• Project End Date: Sep 30, 2015
• Program Code: [(N/A)]- (N/A)

Recipient Organization
UNIVERSITY OF CALIFORNIA, DAVIS
410 MRAK HALL
DAVIS,CA 95616-8671

Performing Department
Population Health & Reproduction

Non Technical Summary
In 2004, the World Health Organization (WHO) recommended zinc supplementation to oral rehydration salt (ORS) for 10-14 days to decrease duration and severity of infant diarrhea.3 The addition was in response to numerous randomized clinical trials which consistently showed benefits to treating acute gastroenteritis with ORS supplemented zinc. Similar benefits of zinc-supplemented ORS on diarrhea in calves may have numerous appealing implications including reduced recovery times, decreased infant mortality, and significant impact on the economics and sustainability of antibiotic use in food animals. Although numerous animal studies report on zinc and its beneficial effects or toxicosis, zinc's therapeutic use in calves with diarrhea is yet to be studied.

Animal Health Component

100%

Research Effort Categories

Basic

(N/A)

Applied

100%

Developmental

(N/A)

Classification

Knowledge Area (KA)	Subject of Investigation (SOI)	Field of Science (FOS)	Percent
311	3410	1170	90%
302	3410	1010	10%

Knowledge Area
302 - Nutrient Utilization in Animals; 311 - Animal Diseases;

Subject Of Investigation
3410 - Dairy cattle, live animal;

Field Of Science
1010 - Nutrition and metabolism; 1170 - Epidemiology;

Keywords

zinc

gastroenteritis

diarrhea

calves

oral rehydration solution

Goals / Objectives
RESEARCH HYPOTHESES. 1) In 1 to 21 day-old Holstein calves with diarrhea, daily oral administration of zinc-supplemented ORS for the period of clinical signs or a maximum of 14 days reduces the severity of disease as evident by improved fecal consistency, higher recovery rates and lower mortality rates. 2) In 1 to 21 day old Holstein calves with diarrhea, daily oral administration of zinc-supplemented ORS for up to 14 days does not result in zinc toxicity or a deficiency of other heavy metals. 3) ORS supplemented with inorganic zinc such as zinc oxide has comparable effect to protein-bound zinc formulations suggesting a local effect in the GI tract. OBJECTIVES. 1) Compare fecal scores, recovery and mortality rates in 0 to 35 day-old Holstein calves (up to 21 days at enrollment + 14 days maximum treatment) with diarrhea randomly assigned to either zinc-supplemented or conventional zinc-deficient ORS daily treatment for a maximum of 14 days or until normal fecal consistency. 2) Investigate zinc toxicity and heavy metal deficiency in a subset of Holstein calves treated orally with zinc-supplemented ORS for 14 days. 3) Compare effectiveness of inorganic versus organic zinc formulations of ORS for treatment of diarrhea in Holstein calves.

Project Methods
PROJECT DETAILS. The study will be conducted on a client herd of the VMTRC with male Holstein calves between 1-21 days old enrolled upon developing diarrhea. Although toxicity or health losses in study calves is highly unlikely, the calves will be purchased upon enrollment yet housed for a daily fee on the source herd to maintain field conditions. A total of 36 calves will be randomly allocated to one of three treatment groups, a group of calves treated twice daily with oral zinc oxide supplemented ORS, a second group treated with zinc methionine and a third group treated daily with oral zinc-deficient ORS. Calves will be block randomized into the three study groups with age as a blocking factor to maintain comparable age distribution among groups. A total daily dose of 80 mg of zinc methionine or zinc oxide will be used to supplement the ORS of the respective study groups and which is based on the ratio of human zinc nutrient requirement30 to zinc supplementation of ORS(1:6.7) and which was verified to be below a calf's ratio of normal zinc intake29 to toxic levels and which was 1:37.5. At enrollment, a liver biopsy and venous blood samples will be collected for heavy metal screening including zinc. Thereafter, fecal and blood samples will be collected every other day for screening. Similarly body weight, serum total protein, and blood packed cell volume will be assessed daily to determine body weight changes and calf hydration. In addition, attitude, body condition score and fecal consistency will be recorded daily. Treatment will continue in all calves until clinical signs resolve or for a maximum of 14 days during which a second liver biopsy will be also obtained to assess treatment. Fecal samples collected at enrollment and end of follow up (last fecal sample prior to recovery, death or censoring) will be tested at CAHFS laboratory using a commercial kit that has been validated for detection of bovine Rota and Corona viruses and Escherichia coli K99 in calves. Study calves lost to follow-up due to death will be necropsied to determine the cause and obtain mean jejunal crypt heights as described elsewhere. Anticipated results. We expect to find a significant reduction in days to recovery, mortality rate, weight loss, minimum dehydration score in calves treated with zinc compared to controls. Current knowledge supports an expectation of similar findings in calves treated with zinc methionine compared to those treated with zinc oxide, however, it is possible that zinc's primary mode of action is local intra-luminal antibacterial, protective mucosal coatant, or an unknwon mechanism and which may result in higher effectiveness in zinc oxide treated group compared to zinc methionine. We also expect that the oral supplementation at the proposed daily dose for 14 days will not result in a toxicity or alter the levels of other heavy metals.

Progress 10/01/11 to 09/30/15

Outputs
Target Audience:The target audience of this study are veterinarians, nutritionistand producers working on calf health, feedig and design of prevention programs, specifically preweaned dairy calves. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Dr. Glover was the Production Medicine Resident at the Veterinary Medicine Teaching and Research Center at the School of Veterinary Medicine University of California Davis. This study served as her Master in Preventive Veterinary Medicine (MPVM) thesis which she acquired through its implementation experience in study design, implementing calf helath studies, literature review, statistical analysis and write up. Dr. Glover won the second place award in the American Association of Bovine Practitioners award for best graduate student presentation for this study. How have the results been disseminated to communities of interest?Through publication, presentation at American Association of Bovine Practitioners and producer meetings including one of the two largest calf ranches in the US. What do you plan to do during the next reporting period to accomplish the goals?We planned on concudting a follow up trial investigating the preventive role of zinc. Which we did (also reported in REEPORT).

Impacts
What was accomplished under these goals? This is the first randomized clinical trial testing theeffect of a daily oral zinc supplementation in neonatal Hol-stein calves with diarrhea. Data collected from the currenttrial did not provide evidence for a beneficial effect of dailyoral 80 mg of zinc as either zinc Met or ZO treatment ondays to recovery, fecal consistency, or microbiological andclinical cures in Holstein calves with diarrhea. A total dailydose of 80 mg of available zinc did not result in a zinctoxicity or mineral deficiency. Although statistically notsignificant, the current trial identified clinically importantdifferences in weight change during the course of diar-rhea in calves and other potentially interesting contrastsbetween effect of organic versus inorganic zinc formula-tions on clinical and microbial cures in neonatal Holsteincalves which differed by pathogen.

Publications

• Type: Journal Articles Status: Published Year Published: 2013 Citation: http://dx.doi.org/10.1016/j.prevetmed.2013.09.001 A double-blind block randomized clinical trial on the effect of zinc as a treatment for diarrhea in neonatal Holstein calves under natural challenge conditions. AD Glover, B Puschner, HA Rossow, TW Lehenbauer, JD Champagne,P.C. Blanchard, S.S. Aly Preventive veterinary medicine 112 (3-4), 338-347

Progress 01/01/13 to 09/30/13

Outputs
Target Audience: Dairy cattle owners managing dairy herds, calf ranches etc. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? Dr. Alex Glover was the graduate student on this project, she was funded for her research and Masters in Preventive Veterinary Medicine degree which she completed. In addition, training for staff at the calf ranch was conducted on best management practices for calf diarrhea and calf euthanasia. Finally, DVM degree students and undergraduate students were trained on liver biopsy for this and other projects that required that technique. How have the results been disseminated to communities of interest? 1. Glover AD, Puschner B, Rossow HA, Lehenbauer TW, Champagne JD, Blanchard PC, Aly SS.A double-blind block randomized clinical trial on the effect of zinc as a treatment for diarrhea in neonatal Holstein calves under natural challenge conditions. Prev Vet Med. 2013 Nov 1;112(3-4):338-47. PubMed PMID: 24074841. 2. Glover, A. D., Puschner, B., Rossow, H. A., Champagne, J. D., Lehenbauer, T. W., Blanchard, P.C., Aly, S. S.A Randomized Clinical Trial for the Effect of Oral Zinc Supplementation as a Treatment for Diarrhea in Neonatal Holstein Calves.Proceedings of the 45th Annual Conf, Am Assoc Bovine Pract (AABP). What do you plan to do during the next reporting period to accomplish the goals? Goals accomplished for this set of objectives, funding pending for subsequent year’s proposal.

Impacts
What was accomplished under these goals? 1) In 1 to 21 day-old Holstein calves with diarrhea, daily oral administration of zinc supplemented ORS for the period of clinical signs or a maximum of 14 days reduces the severity of disease as evident by improved fecal consistency, higher recovery rates and lower mortality rates. 2) In 1 to 21 day old Holstein calves with diarrhea, daily oral dministration of zinc-supplemented ORS for up to 14 days does not result in zinc toxicity or a deficiency of other heavy metals. 3) ORS supplemented with inorganic zinc such as zinc oxide has comparable effect to protein-bound zinc formulations suggesting a local effect in the GI tract. OBJECTIVES. 1) Compare fecal scores, recovery and mortality rates in 0 to 35 day-old Holstein calves (up to 21 days at enrollment + 14 days maximum treatment) with diarrhea randomly assigned to either zinc-supplemented or conventional zinc-deficient ORS daily treatment for a maximum of 14 days or until normal fecal consistency. 2) Investigate zinc toxicity and heavy metal deficiency in a subset of Holstein calves treated orally with zinc-supplemented ORS for 14 days. 3) Compare effectiveness of inorganic versus organic zinc formulations of ORS for treatment of diarrhea in Holstein calves.

Publications

• Type: Journal Articles Status: Published Year Published: 2013 Citation: Glover AD, Puschner B, Rossow HA, Lehenbauer TW, Champagne JD, Blanchard PC, Aly SS. A double-blind block randomized clinical trial on the effect of zinc as a treatment for diarrhea in neonatal Holstein calves under natural challenge conditions. Prev Vet Med. 2013 Nov 1;112(3-4):338-47. PubMed PMID: 24074841.

Progress 01/01/12 to 12/31/12

Outputs
OUTPUTS: Results of this research were presented in AABP 2012 in Canada and as a thesis defense for the MPVM program in December of 2012. In addition the manuscript has been submitted to JDS. PARTICIPANTS: Coauthors collaborated to complete the project testing, analysis and write up for publication. TARGET AUDIENCES: Private practitioners. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.

Impacts
Diarrhea is the leading cause of death in neonatal calves and contributes to major economic losses. The objective of this randomized clinical trial was to evaluate the effect of oral inorganic and organic zinc supplementation as a treatment for neonatal Holstein calves with diarrhea. Seventy nine 1 to 8 day old male Holstein calves on a California calf ranch located in Tulare County were block randomized to one of 3 treatments within 24 hours from their first onset of diarrhea after birth. Calves received a daily dose of either a placebo composed of 80 mg of zinc-free powder, 381.54 mg of zinc methionine (ZM) (equivalent to 80mg of zinc), or 99.69 mg of zinc oxide (ZO) (equivalent to 80 mg of zinc) in 2 L of a zinc-free oral rehydration solution (ORS). Calves were treated once daily until normal fecal consistency or for a maximum of 14 days. Upon enrollment and exit, calves were weighed, and blood, feces, and liver biopsies were collected for trace mineral analysis. Fecal samples at enrollment and exit were tested for E. coli K99, Cryptosporidium spp., rotavirus and coronavirus. Baseline liver zinc concentrations for the 71 calves in the placebo, ZM, and ZO treatment groups were 710.6 (SEM = 147.7), 852.3 (SEM = 129.6), and 750.7 (SEM = 202.9) mg/kg dry weight (DW) respectively. A non-significant trend in calves supplemented with ZM for a mean weight gain of 40 grams/d compared to a mean weight loss of 67 grams in the placebo group calves. Cryptosporidium-positive calves treated with ZM had higher odds of testing negative at exit compared to placebo calves (Odds Ratio, OR = 12.0). Although not statistically significant, calves supplemented with ZO tended to recover one day earlier compared to calves treated with a placebo (8.5 d vs. 9.7 d). Similarly, calves that tested positive for coronavirus at enrollment and treated with ZO had a higher odds of clinical cure compared to calves treated with a placebo (OR =3.14). Data collected from the current trial did not provide evidence for a beneficial effect of zinc on days to recovery, fecal consistency, or microbial and clinical cure in Holstein calves with diarrhea. However, further research on the use of zinc to mitigate the adverse effects of diarrhea in calves is warranted.

Publications

• No publications reported this period