All information in this article is for educational purposes only.  It is not for the diagnosis, treatment, prescription or cure of any disease or health condition.

 

 

AHVMA Journal ¥ Volume 48 Fall 2017

 

TISSUE MINERAL ANALYSIS PATTERNS

IN 564 DOGS

By Ava Frick, DVM and Arlene Tolen, MPH, NC

 

ABBREVIATIONS

ARL – Analytical Research Laboratory

CLIA – Clinical Laboratory Improvement Amendments

TMA - Tissue Mineral Analysis

Note: Standard elemental abbreviations are used throughout, for example Mg for Magnesium

 

Abstract

Tissue Mineral Analysis (TMA) is a technique using soft

tissue hair or fur biopsy that provides a reading of the

mineral deposition in the cells and interstitial spaces of

the hair over a 2 to 3 month period. TMA can be used to

understand metabolism. It is another scientific measure that

can expand our understanding of health and processes that

impact illness in dogs. Mineral excess or deficiency is known

to produce certain physical and psychological symptoms.

The correlation of TMA results with clinical signs seen in

patients is discussed in this paper.

Tissue mineral levels and electrolyte patterns of calcium

(Ca), magnesium (Mg), sodium (Na), and potassium (K)

were analyzed in 564 dogs (300 male, 264 female; 99%

neutered or spayed) of variable breeds. Their ages ranged

from 1 to 15 years. Cases included all dogs presented to the

authors within a 12-year period.

Ninety-four percent (94%) of the 564 dogs were found to

have high Na to K ratios with low Ca and Mg. Ca, Mg, Na,

and K are prevalent minerals in the body, regulate osmotic

balance, and are involved in most body functions. According

to mineral analysis research, both results (levels and ratios)

are indicative of inflammation.

TMA is an accurate and scientifically valid approach to

assessing metabolic function and balance. TMA is responsive

not only to the trace mineral levels in the diet, but also to

all other factors which influence their metabolism including

stress, exercise, and endocrine and gastro-intestinal function.

 

INTRODUCTION

Hair mineral analysis is a tissue mineral biopsy blueprint of

oneÕs biochemistry and the nutritional metabolic activity

that occurred during the period of hair formation. Hair

and fur are chemically indistinguishable, having the same

chemical composition, and are made of keratin. The primary

difference between hair and fur is the word usage. The hair

of non-human mammals is referred to as Òfur,Ó while humans

are said to have hair. For the purposes of this paper, ÒhairÓ

refers to both hair and fur.

Hair is formed from clusters of matrix cells that make up

the follicles. During the growth phase, the hair is exposed to

the internal metabolic environment such as the circulating

blood, lymph, and extracellular fluids. As hair continues

to grow and reaches the surface of the skin, its outer layers

harden, locking in the metabolic products accumulated

during the period of formation. Hair analysis reveals

individual metabolic states in addition to the mineral status

of the individual animals (1, 2, 3).

Hair is 10-15% porous, and washing agents used in the

laboratory process can remove not only the exogenous

elements but also penetrate inside the shaft and wash

out some of the loosely bound minerals. Washing a

sample during test preparation removes quantities of

water-soluble macrominerals. When hair samples are not

washed by the laboratory prior to assessment, the analysis

can accurately measure macrominerals. Therefore, it

is important to select a laboratory that does not wash

the submitted specimen prior to the mass spectrometer

analysis.

The laboratory will instruct the practitioner on the

protocol for collecting the sample from the animal (4-7).

For animals treated with topical insecticides and pesticides, it

is recommended to collect the sample during the last week of

the treatment cycle (i.e., 1 week before the next application

for most products). The animal can be bathed and clipped 4

hours later. Alternately, the ventrum, where the sample will

be collected, can be lightly sprayed with alcohol, wiped clean,

and then clipped, avoiding the topical application zone.

TMA is a non-invasive and a cost-efficient screening test (8).

It has been used throughout the world to assess individual or

herd nutritional status, providing a sensitive indicator of the

long-term metabolic trends from the effects of diet, stress, and

toxic metal exposure (9-14). Most deficiencies in animals are

brought about by altered relationships of minerals within the

body. It has become evident that both the retention and loss of

minerals by the animal are equally as important as the nutrients

consumed from the feed itself. This is valuable in determining

dietary needs as well as for recommending supplementation

(15-17).

Mineral ratios and levels can be changed by the

presence of toxic metals, nutritional deficiencies, infections,

illness, or stress from a myriad of sources (18, 19).

Assessment of an individualÕs stage of stress and oxidation

rate (how fast the body metabolizes the food in exchange for

energy) can be determined by TMA (20). Measured stress

can come from external or internal sources. Helping or

treating stress with specific minerals can improve the health

status, resulting in better physical and emotional responses.

Many factors can influence the stress patterns of a TMA

report.

Trends for common psychological conditions such

as depression, hyperactivity, anxiety, and mood swings may

be identified through TMA utilizing the levels and ratios of

calcium (Ca), magnesium (Mg), sodium (Na), and potassium

(K). Research confirms the intimate connection between

the bodyÕs biochemistry and many emotional states (21-24).

Changes in body chemistry are reflected in the cellular

metabolism as measured by the hair tissue mineral

levels and inter-relationships found via TMA.

These can indicate metabolic dysfunction before clinical signs

occur. While blood values reflect what is in the blood,

hair analysis provides a record of how the body stores

and disposes of elements. The choice of hair as a testing

medium is based on the fact that blood chemistries

change dynamically from day to day, while hair values give

a more stable view of the overall mineral nutrition (25).

The purpose of this retrospective study was to show how

TMA has been used on a large dog population to establish

typical mineral patterns; to present why TMA is a valid test to

aid in nutritional assessment; to ascertain how these patterns

affect function, health and behavior; and to help determine

reasons why this can be occurring in dogs across the U.S.

 

MATERIALS AND METHODS

 

Tissue mineral levels and electrolyte patterns of Ca, Mg, Na

and K were analyzed in 564 dogs (300 male, 264 female; 99%

neutered or spayed) of variable breeds. Their ages ranged

from 1 to 15 years and included all dogs presented to the

authors within a 12- year period. The health conditions or

clinical signs of the dogs ranged from dermatitis, anxiety,

nervousness, aggression, gastrointestinal conditions,

metabolic conditions, neuromuscular, pain, and degenerative

joint disease. Some samples from apparently healthy dogs

were run for a baseline assessment by their owners (Table 1).

Table 1: Presenting Clinical Signs or Conditions

Presenting Clinical Signs or Conditions Number of Dogs

Included in Group

Dermatological 370

Anxiety, Aggression, Nervousness 302

Gastrointestinal 252

Metabolic Conditions (diabetes, heart disease, liver

disease, kidney disease, cancer)

218

Pain 201

Neuromuscular 151

Degenerative Joint Disease 117

Apparently Normal 47

 

Seven groups of conditions or clinical signs that were seen in 517 of the total

group are listed here. Many of the dogs had multiple problems.

Another 47 dogs were apparently healthy without any complaints

from the owners. The TMA in those 47 was run as a baseline.

Their data is also included in the total statistics reported.

The fur was collected via clipping or cutting from the ventrum,

not longer than 1.25 cm in length and taken close to the skin.

Samples were weighed on a small scale provided by the laboratory

to at least 100 mg (less than 100 mg would produce insufficient

sampling), placed in a paper envelope, and submitted to the

Analytical Research Laboratory (ARL) in Phoenix, Arizona.

Computer-controlled mass spectrometers and induction-coupled

plasma (ICP) instruments are used today at hair testing

laboratories in the U.S. All commercial hair testing laboratories

in the U.S. are licensed and inspected annually by the federal

government as part of the Clinical Laboratory Improvement

Amendments (CLIA) act.

In this study, testing was performed

on a Perkin Elmer Elan 9000 ICP Mass Spectrometer. Results

are reported as mg % (parts per million times 10). The source

of standards for TMA used by the ARL is provided from original

dog research by Parmae Laboratory (Texas) along with 40 years

of research data collected by Dr. Paul Eck of ARL. A completed

report includes the optimal reference range, the patientÕs level,

toxic metals, and mineral ratios (Table 2).

((sTable 2: Example of TMA report

Discussion

 

There are thousands of biochemical reactions that ultimately

control metabolism, digestion, and the regeneration of

body tissues. The vast majority of these reactions depend on

minute amounts of trace minerals for their activity. If these

essential minerals are not present to fuel the processes, then

the bodyÕs ability to regenerate, metabolize, or break down

noxious substances is compromised.

Because hair in the growth phase is exposed to the internal

metabolic environment (circulating blood, lymph, and

extracellular fluids) and retains the metabolic products

presented to it as it hardens, it becomes a perfect tissue sample

for testing body function, metabolic trends, and toxic metals.

TMA is a technique using soft tissue mineral biopsy that provides

a reading of the mineral deposition in the cells and interstitial

spaces of the hair over a 2 to 3 month period. This form of

testing allows for measurement capabilities of nutritional

alterations and improved recommendations for vitamin

and mineral supplementation or nutrient augmentation.

Ninety-four percent (94%) of the 564 dogs in the study

population had low Ca, Mg, and K, and high Na. This

correlates to a metabolic stress pattern, and the consistency of

these mineral patterns should not be ignored. Other studies

have reported similar findings; they linked this mineral

pattern to inflammation influencing T cells, cytokines, and

anti-inflammatory plasma proteins (26, 27). Another set

of parallel data confirmed that the deficiency of essential

mineral elements and Na overload can directly cause lipid

peroxidation and eventually hepatic damage (28).

Hair analysis offers a perspective for studying both

ÒdiseaseÓ trends and for studying ÒstressÓ and physiological

coping mechanisms (29, 30). Disease trends and metabolic

changes can be documented and supported by using hair

TMA (31-33).

Ca and Mg levels have been measured in patients with

fibromyalgia (34). Hair Ca concentration also correlates

to coronary heart disease (35). Levels of Ca and Mg have

been studied as part of the metabolic syndrome (36). Both a

deficiency and excess of these electrolytes have been shown

to be of pathogenic value in the development of endocrine

disorders and thyroid disease (37-40).

Nutrients can be unavailable for many reasons.

Malabsorption or bio-unavailability of specific minerals

can occur due to gut interference; improper form; lack

of other needed vitamins, minerals, or amino acids; and

ligand interruption, to name a few. Low taurine, low K,

and high Na all lead to low tissue Mg levels. Studies of both

animals and humans link minerals to healthy functioning

of the nervous system and behavior (41, 42). Nutrient

minerals must be available to support positive behavior.

Ca plays a fundamental role in metabolic processes, which

can be altered by small changes in extracellular ionized

Ca concentrations (43-46). A tissue Ca deficiency can

exhibit as a sympathetic dominant state (alarm or frightflight

reactions), anxiety, bruising, high blood pressure,

ÒfastÓ oxidation/metabolic state, insomnia, irritability,

muscle cramps and spasms, nervousness, hyperactivity,

osteoporosis, weakened ligaments and tendons, and tooth

decay (47,48). When the extracellular fluid level of Ca

ions drops below normal, the nervous system becomes

progressively more excitable because of increased

permeability of the neuronal membrane to Na (49).

A deficiency of tissue Mg affects the autonomic nervous

system, behavior, muscle tone and function (50-53). Clinical

signs include poor appetite, irritability, weakness, muscle

tremors, tetany, twitching, numbness, tingling, confusion,

disorientation, personality changes, apathy, memory loss,

skin lesions, tissue calcification, elevated cholesterol,

cardiovascular changes, tachycardia, elevated parathyroid

hormone, pancreatitis, and stress (42).

Magnesium deficiency increases catecholamine

secretion and sensitivity to stress, and may promote

aggressive behavior (54). Overall, Mg can act as a

neuron protector against aggression. Magnesium helps

to protect the nervous system locally and globally, is a

psycho-stabilizer, and plays an important role in aging

(55, 57). Increased catecholamine-induced intracellular

Mg loss is a causal factor in urinary loss of Mg (56).

When tissue K is deficient relative to Na, the body will tend

to develop hypertension, arrhythmias, and fatigue (57).

Excess Na will be able to enter the cell, which interferes with

cellular metabolism, especially protein synthesis. When this

happens, the cellular response is expressed as loss of energy

and manifests in the individual as exhaustion, submissive

behavior, and heightened anxiety or panic (58).

High tissue Na levels are indicative of excessive adrenal

gland activity and excitability. Excess Na combined with

too much phosphorus, low tissue levels of Ca, Mg, zinc

(Zn), and K result in adrenal insufficiency (59). Na,

when elevated in the tissue, is a stress that is associated

with anger and fear, the emotions of the Òfight or flightÓ

response, and an indication of body inflammation (60).

With the activation of the stress response, the adrenal glands

increase their secretion of the hormone aldosterone, which

increases the retention of Na in the cells and tissues. As Na is

retained, Mg and Ca are lost from the cells and tissues. The

loss of Zn and Mg under stress appears to facilitate the stress

response by allowing for greater retention of Na (61, 62).

The primary mechanism by which psychological factors

predispose one towards a disease process is by means of the

stress response and its effect on nutrient minerals at the cellular

level. The stress response involves general systems responses

in which psychological, neurological, endocrine, and immune

system phenomena occur. Cellular energy production and

neuro-endocrine interference ensues. Mineral imbalance is a

factor in physiological stress and antisocial behavior (63, 64).

Stress has the capacity to impact minerals even more than

other nutrients. Zn and Mg are lost from the bodyÕs tissue

reservoir in the ÒAcute StageÓ of stress. Ca and copper are

lost in the ÒResistance StageÓ of stress. Loss of Na and K

are associated with the ÒExhaustion StageÓ of stress. Both

internal and external stressors create an increased need for

vitamins and minerals to recover and to prevent depletion

and breakdown.

Dogs are carnivorous biased, which means they derive

optimal health and well-being by eating meat. Even though

they have omnivorous capacity and many manage to survive

on grain and carbohydrate-rich foods, a solely grainbased

diet is not the best choice for maximizing digestion,

absorption, health, and disease-free longevity.

Grains have been shown to interfere with Zn and

Ca assimilation (46). Eating carbohydrates lowers

Ca and Mg, and raises Na. Ca and Mg are sedative

elements, or known as the calming minerals. Lowering

their levels therefore increases the sensitivity of the

sympathetic division of the autonomic nervous system

and prepares the dog to respond to either fight or flight.

Mineral composition of the body is dependent not only

on food intake but on the efficiency or inefficiency of

neuroendocrine function (65). Hormones are known to

influence nutrients at several levels including absorption,

excretion, transportation, and storage. Nutrients in

turn can exert an influence on hormones (39, 66, 67).

The generations of dogs who were fed the dry grain-based

foods may have an impact on todayÕs canine

offspring that are now showing physiological imbalance.

Pesticides, herbicides, and insecticides are known to act as

endocrine disruptors in humans and animals. Herbicides

and fungicides affect Ca transport in plant mitochondria.

Herbicides and insecticides have been shown to interfere

with Ca metabolism in birds. Further, 354 drugs have been

reported to interact negatively with Ca, Mg, and Zn. Toxic

metals affect mineral metabolism and availability. Tannins

and polyphenols from tea, for example, reduce Mg in

tissue. These are all stressors that affect minerals (68, 69).

 

Conclusion

 

The majority (94%) of the 564 dogs studied were found to

have a high Na/K ratio with low Ca and Mg. According to

mineral analysis research, both results (the levels and the

ratio) are indicative of inflammatory processes and stress. This

correlates well with the clinical signs (general, physiological,

and psychological) reported in the cases studied.

Macrominerals are essential in metabolism in that they

activate catalytic and enzyme functions. Hair analysis is used

not only for measuring those minerals but also to monitor

the nutritional state of the dog until treatment benefits are

achieved and the effects of the treatment protocol have been

stabilized. The combination of feed ratio and hair analysis is an

invaluable screening tool to determine the correct program of

diet and supplementation for each dog's specific needs. TMA

is responsive not only to trace mineral levels in the diet but to

all other factors which influence their metabolism including

stress, exercise, and endocrine and gastro-intestinal function.

When performed to standards and correctly interpreted,

a TMA can be used as a screening tool in canine wellness

programs; and for those in suboptimal health, for monitoring

mineral deficiencies, mineral excesses, biochemical

characteristics, system imbalances, supporting behavior,

and endocrine indices (70-75). The use of hair TMA offers

a sophisticated and cost-effective approach (less than a

typical chemistry/CBC panel) that is very affordable for any

practitioner to incorporate into their nutritional assessment

evaluations.

There is currently nothing else available in the

way of a simple laboratory test that gives the veterinarian or

nutritional consultant a perspective of the internal nutritional

status of an animal. It can be a method to comply with AVMA

and AAHA recommendations regarding the importance

of offering annual or semi-annual nutritional evaluations.

This enables the veterinarian to present a nutritional

consultation that has significant value and direction toward

resolving health issues using individualized nutrition.

 

Acknowledgment

 

Technical assistance was recruited from Elaine Eisenbeisz of

Omega Statistics in Murrietta, California for the statistical

analysis. Manuscript submission preparation was enlisted

from Lisa Winstead in Rolla, Missouri.

 

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