Effect of a Commercially Available Electrolyte Solution on Acute, Non-Specific Diarrhea in Dogs ()
1. Introduction
Dogs housed in animal shelters frequently developed non-specific diarrhea that is commonly associated with bacteria, viruses, or parasites [1] . Nematodes like Ancylostoma caninum, Toxocara canis, and Trichuris vulpis and the protozoans Giardia lamblia, Cryptosporidium canis, and Cystoisospora spp. are commonly found to be involved in non-specific diarrhea cases [2] . Dehydration and electrolyte disturbances are common in cases of diarrhea and the management of these problems is a large part of many budgets in shelters. In addition, diarrhea causes discomfort and slows adoption which also increases expenses to shelters. Due to negative effects on the fecal microbiome, current recommendations suggest not using oral antibiotics routinely in dogs with non-specific diarrhea. Thus, other treatment pathways for the management of acute non-specific diarrhea are needed [3] .
The electrolyte solution (ES; SxDog, EcoPlanet), is classified as a nutritional supplement that has been shown to be effective in the management of dehydration and diarrhea in calves (https://ecoplanetonehealth.com/). The inventors have shown safety in a limited number of dogs in an unpublished open trial when used to treat non-specific diarrhea in several local veterinary clinics.
The primary objective of Experiment 1 was to assess whether the product is safe to administer to healthy beagles and to determine the effects on the fecal microbiome. The hypothesis for Experiment 1 was that ES would have no noticeable side effects or toxicity and would have no negative effects on the fecal microbiome.
The primary objective of Experiment 2 was to dose dogs with diarrhea in a shelter the ES with a prescription diet, the ES with a standard diet, or a prescription diet alone. The hypothesis for Experiment 2 was that diarrhea response rates would be similar amongst the groups of dogs.
2. Methods
2.1. Experiment 1—Healthy Dogs
A total of six healthy laboratory raised mixed sex, 2- to 5-year-old beagles were used in this IACUC-approved study (170-052). The dogs were on a standard facility diet for over 12 weeks and were housed individually.
On Day 0, physical examinations were performed, and blood was collected for complete blood cell counts (CBC) and serum biochemical panels. In addition, feces were collected for fecal flotation, Giardia and Cryptosporidium immunofluorescent antibody assay (MeriFluor, Meridian Diagnostics), and for fecal microbiome assessment (Gastrointestinal Laboratory, Texas A & M University, School of Veterinary Medicine & Biomedical Sciences, College Station, TX). Dogs were then treated with the ES at 1.5 ml/kg on Day 0 and Day 1. The dogs were evaluated daily on Days 0-14 by abdominal palpation and for attitude, appetite, vomiting, diarrhea, inappetence, and for a fecal character using a standardized scoring system (Table 1). Tympanic membrane temperatures were recorded daily. CBC and serum biochemical panels were repeated on Days 7 and 14 to evaluate for toxicity.
2.2. Experiment 2—Clinical Trial
Dogs housed in a shelter in north-central Colorado with acute diarrhea (score of >4) were approved for entry by the shelter veterinarian. Dogs with inappetence, fever, BW loss > 2%, dehydration > 3% - 5%, vomiting, abdominal pain, abdominal masses, or a positive parvovirus antigen test on feces were excluded.
Feces from all dogs were tested on site at the shelter for Giardia spp. using a commercially available antigen assay (SNAP Giardia; IDEXX Laboratories, Portland ME). Feces were also evaluated by fecal flotation and Giardia/Cryptosporidium IFA. After the SNAP Giardia assay was performed, 2 grams of feces
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Table 1. Clinical scoring parameters were measured daily by masked, trained observers in Experiment 1 and the fecal scoring rubric used in both Experiments 1 and 2.
were transferred to the Center for Companion Animal Studies for microscopic examination for parasite eggs, cysts, and oocysts after sugar centrifugation as well as a commercially available fluorescent antibody assay for Giardia cysts and Cryptosporidium oocysts (Meridian Diagnostics). Dogs positive for Giardia spp. had the assemblage determined as previously described [4] . The fecal flotation and fluorescent antibody assay were performed again on Day 5 or the day the dog was adopted.
A random number generator was used to assign the treatment groups which included the ES with the facility diet, the ES with a veterinary prescription diet (Hill’s Pet Nutrition Prescription Diet i/d), or the veterinary prescription diet alone. Dogs with Giardia or nematodes were administered fenbendazole at 50 mg/kg, PO daily for five days. The feces were scored daily (Table 1). The body weight of each dog was determined on Day 0 and Day 5 or the day the dog was adopted. A medicine consultation was available for these cases as requested by the shelter veterinarian.
3. Statistical Evaluation
To be included in the final data analysis, each dog was enrolled in the study for at least 4 days. The proportion of dogs with parasites by groups were compared by Fisher’s exact test. The mean and standard deviations for the days to the first normal stool were calculated for each group and compared by student’s t-test. Significance was defined as P < 0.05 in all analyses.
4. Results
4.1. Experiment 1—Healthy Dogs
All six dogs were administered the protocol approved dose of ES on two consecutive days (1.5 ml/kg, PO). There was no salivation or vomiting noted in any of the dogs during or after administration. There were no significant findings in any of the four monitoring rubrics (Table 1). The body weight changes for each dog over the course of the study were within normal limits. A total of five dogs had complete data sets and no significant changes were noted on the CBC. One dog had a clot in the pre-treatment complete blood cell count sample, but the Day 7 and Day 14 results were all within normal limits. Complete data sets were available for all dogs and there were no changes of significance over time that could relate to toxicity. In the microbiome assessment of the healthy dogs, there was no evidence of a dysbiosis.
4.2. Experiment 2—Clinical Trial
A total of 43 dogs were screened for inclusion in the study. Of these dogs, a total of 22 dogs (10 dogs fed ES plus i/d; seven dogs fed placebo and i/d alone; five dogs fed ES and a standard diet) remained in the study long enough to generate data for analysis. Parasites detected included Giardia spp., Cystoisospora spp. Cryptosporidium spp., Ancylostoma caninum, and Toxocara canis (Table 2).
The mean and standard deviation of the day to first normal stool after the first dose of ES or placebo was calculated for the three groups of dogs and no significant differences were recognized (Figure 1). Overall, 16 of the 22 dogs had a normal stool the day after the first dose of ES or placebo. Of the six dogs with a slow response to normalcy (first normal stool detected after Day 1), two dogs were fed ES plus i/d, one dog was fed placebo and i/d alone, and three dogs were fed ES and a standard diet.
All six dogs with the first day of normal stool detected after Day 1 were parasitized and when the mean and SD of the days to normalcy were compared between parasitized dogs (1.7 ± 0.99) and dogs with no parasites (1 ± 0), the parasitized dogs had a significantly slower response (P = 0.018). All six dogs with a coccidian parasite (Cryptosporidium spp. or Cystoisospora spp.) were in an ES group and of these dogs, four had a response to normalcy > 1 day. Of the three
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Table 2. Distribution of parasites in the dogs of the study. There are no significant differences in the prevalence rates amongst the three groups (P > 0.05).
a. ES = Electrolyte Solution. b. i/d = Hill’s Pet Nutrition Prescription Diet i/d.
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Figure 1. The number of days to normalcy for each of the three groups: ES and i/d, placebo and i/d, and ES and normal diet.
dogs with a response to normalcy on Day 3 or Day 4, all were infected with Cryptosporidium spp. or Cystoisospora spp. There were six dogs infected with Giardia spp. alone and each of these dogs was normal by Day 1 (four dogs) or Day 2 (two dogs). All the Giardia positive dogs with adequate DNA for sequencing were infected by the non-zoonotic assemblage C genotype.
5. Discussion
In Experiment 1, the ES was shown to be safe in laboratory reared dogs. This conclusion was based on clinical findings (no vomiting or diarrhea) and no abnormal biochemical findings. Additionally, there was no induction of dysbiosis detected from the microbiome analysis.
In Experiment 2 in shelter dogs, it was shown that ES could be used with whatever diet they were currently being fed. The results of this study suggest that ES added to a standard diet is equivalent to using a therapeutic diet alone or the ES combined with a therapeutic diet. However, the major limitation is the sample size. Not having to use a therapeutic prescription diet potentially saves money and the time it might take to get a prescription diet ordered for owners.
In this study, it was shown that parasitism, particularly in the coccidian group, may lead to a slower response and resolution of diarrhea than a viral or bacterial agent. Therefore, the results support doing a fecal examination in the workup of shelter dogs with acute diarrhea. In a future study, it would be interesting to see if clinical responses were improved if the electrolyte solution was combined with specific drugs for the parasite that was diagnosed. Finally, the Giardia spp. identified from the fecal samples were not zoonotic, which is similar to many studies performed in the USA in general and the USA in specific [5] [6] .
Acknowledgements
The authors thank Katrina Jones and Megan Slaughter for their help during data collection and Animal Friends Alliance for the use of their facilities. Funded by unrestricted donations to the Center for Companion Animal Studies (Web address: http://csu-cvmbs.colostate.edu/vth/veterinarians/research/companion-animals/Pages/default.aspx).