Tuscan Red Life Winter Edition 2023 - English version

Tuscan RedLife

Winter 2023, Issue 4 - English edition

CARE FOR YOUR IRISH TERRIER

SPECIAL HEALTH ISSUE

GENETIC TERMS P

COMMON HEALTH RISKS

CYSTINURIA

P 17

MEET STEFANO CHIESI

The first two pages of this issue are dedicated to those Tuscan Red dogs waiting for their loved ones on the road to rainbow bridge:

Sujoncla Ghost Dance (2007-2015)

Tuscan Red Lucente (2018-2020)

Tuscan Red Brunello Biondi Santi (2011-2021)

Tuscan Red Sassicaia (2011-2022)

Tuscan Red Brunello Bramante (2011-2022)

Tuscan Red Galatrona (2015-2022)

P 7 Genetics Glossary

P 17 Cystinuria

P 5, 39, 40 Cucciolata Z

Welcome

Welcome to this fourth edition of the Tuscan Red newsletter. The scope of this project is to share the activities of the kennel with the Tuscan Red community and to formally introduce those people who make possible future generations of this rare breed of dog: both the families of Tuscan Red and my colleagues across the world. This issue has a principle focus, the disorder known as cystinuria.

All the families who accept puppies or adults from Tuscan Red into their home agree to keep them available as participants in the breeding program that I am constantly researching and building. Our aim is to bring forward new generations of Irish Terriers that conserve the phenotype described in the standard which remains essentially unaltered since its publication in 1876 (In the first stud book prepared by the Kennnel club) and the characteristics of balanced temperament, effervescent and loving that make the Irish Terrier an ideal companion. In this sense I am proud to consider myself a ‘preservation breeder’, a breeder whose objective is to be a custodian of the breed. To this end I adopt as my personal motto the words of the pledge of FCI judges:

Integrity, Study, Transparency

Anne M.Tureen

Litter Z

Tuscan Red Summus and Tuscan Red Ornellaia are happy to announce the birth of the Z litter, 5 males and 6 females, on 16 Feburary 2023.

Summus is Jr Champion and is also expecting a litter in Finland soon! Currently he is preparing for his debut at Crufts 2023.

For this litter we opened the choice of names to all Italian wines, not just Tuscany:

MaschiZephyro,Zenato, Zibbo

Zovetto, Zinzasco.

FemmineZiggurat, Zòjosu, Zaglia

Zisola, Zabaldà, Zeni.

Tuscan Red Summus

Tuscan Red Ornellaia

Keeping Informed!

This is a preparatory section designed to facilitate deeper understanding of the following article. When reading, all pink text is vocabulary defined in the UC Davis glossary provided here. Green text is defined or illustrated in the side columns of the article. For a quick read, follow just the highlighted text.

Quick review of our subject:

Cystinuria is a rare inherited condition. The amino acid Cystine which is normally absorbed by the kidneys is not absorbed in affected dogs. The resulting high concentration of this acid can form crystals and then stones. Due to the narrowness of the urethra, the stones can cause serious blockage of the urinal tract in males. Ultrasound imaging will help diagnose the composition of the stone and subsequent treatment. Surgical removal of the blockage is usually necessary, followed by a low PH diet for the life of the dog. Castration is an important step toward a definitive solution, especially if the condition is linked genetically with the male hormone. This is usually the case in Irish terriers which are classified category III type Cystinuria.

There are currently no tests to determine if an Irish Terrier is genetically predisposed to develop Cystinuria. While tests are available to determine the levels of Cystine in urine, they are not reliable due to the variation in concentration of these acids. Owners of males should monitor the urination of their dogs especially between the ages of 2-6 years, and call the vet immediately if there is blood in the urine, if the dog strains to urinate or there is very scarce urination or no urination at all.

If you are curious to know more about your dog’s inherited characteristics, one of the principal genetic laboratories in Italy, Vetogene, offers a bargain package of 240 genetic tests in a project called My Dog DNA. The long list includes one we are familiar with, Hereditary Plantar (foot) Hyperkeratosis (HFH), and Cystinuria types I A and II A. Many specific conditions are included such as degeneration of the cones of the eye found in Alaskan Malamutes, but also some more common things such as Peripheral sensory neuropathy. Also included are genetic tests relative to the phenotype of the dog, such as eye and hair color, or characteristics such as adaptability to high altitudes. For only 125 euros, certainly an interesting offer.

Genetics glossary

The following is a list of basic terms useful to a general understanding of genetics. The words in pink are used in the following article on Cystinura. Underlined words are defined in this list. Material used By kind permission of University of California Davis Veterinary Genetic Laboratory

• Allele: An allele is an alternate version of a gene This is also known as a variant of a gene. They are caused by differences in the sequence of the DNA at a specific position within a gene.

• Amino acid: Organic molecules that are the building blocks of proteins. There are 20 standard amino acids specified by the information (codon) stored in the DNA

• Autosomal: Of or relating to the autosomes, which are the non-sex chromosomes. In other words, any other chromosomes besides X and Y (in mammals).

• Carrier: An organism possessing one copy (heterozygous) of a recessive allele. The term “carrier” is often used in the context of disease-associated or otherwise deleterious variants. Because carriers are heterozygous, they do not express the phenotype of the recessive allele. However, carriers can transmit the recessive allele to their offspring and can thus produce affected offspring if their mate also contributes a recessive allele.

• Chimerism: When an organism or tissue contains at least two different complete diploid sets of DNA that originated from more than one zygote. In animals this can happen when two fertilized eggs fuse early during gestation or when twins exchange cells.

• Chromosome: Physical structure that stores the genetic information, composed of long strands of DNA coiled around proteins.

• Co-dominance: Both alleles at a locus are expressed equally in an organism. This can lead to the appearance of a “blended” phenotype, but differs from incomplete dominance

o Example: Blood type in humans; a person with blood type AB expresses both A and B proteins on the surface of their red blood cells.

• Codon: A sequence of three consecutive nucleotides in DNA or RNA that specifies the synthesis of a single amino acid (or acts as a start or stop signal) during the translation of mRNA to protein. There are 64 codons, of which 61 specify amino acids and 3 are stop signals.

• Complete penetrance: Traits with complete penetrance will be expressed in all individuals possessing the causal allele

• Compound Heterozygote: A compound heterozygote has two different mutant alleles at the same genetic locus (one on each chromosome).

o Example: There are two variants associated with the tiger eye mutation in Puerto Rico Paso Fino horses, TE1 and TE2. The normal wild type variant is N. A horse can be N/N (no tiger eye), N/TE1 or N/TE2 (no tiger eye,

heterozygous carriers), TE1/TE1 or TE2/TE2 (tiger eye, homozygous recessive), or TE1/ TE2 (tiger eye, compound heterozygote).

• De novo: De novo variants are genetic variations that arise in a reproductive cell (i.e., egg or sperm) or the fertilized egg so that they are present in an individual animal but absent from both parents’ DNA.

• Deletion: A type of mutation in which part of the DNA sequence is lost; the loss can be as small as a single base pair or as much as an entire section of a chromosome

• Dilution (coat color): Dilutions are caused when a variant of a pigmentation gene lightens the coat color of an animal. Dilution mutations can dilute all pigments (both eumelanin and phaeomelanin) or only one pigment type (eumelanin or phaeomelanin). The physical cause of the lightened coat color may vary, from reduction in the amount of pigment produced to inefficient transfer of the pigment from the melanocyte to the hair follicle cells.

o Examples:

 In cats, the dilute allele d creates a dilution of all pigments (and thus all coat colors) by causing the clumping and uneven distribution of pigment granules in the hair shaft.

 In dogs, the Intensity dilution allele In causes an extreme dilution of phaeomelanin (red or yellow pigment), resulting in a cream to white coat in dogs. Eumelanin is not affected.

 In horses, the dun allele D affects both eumelanin (black) and phaeomelanin (red) pigment, diluting the coat but leaving the head, lower legs, mane and tail undiluted.

• Diploid: Having two complete sets of chromosomes; one set is inherited from the mother, and one set from the father.

• DNA: Deoxyribonucleic Acid, a complex organic molecule that is capable of self-replication and that carries and stores genetic information. DNA is the material of heredity and is found in all known living organisms.

• DNA sequence: The ordered sequence of nucleotides on a strand of DNA. Example: TAAGTCTGACACAAT

• Dominant: A trait is dominant when only one copy of a variant allele is needed for an animal to express the phenotype.

• Duplication: A type of mutation in which a gene or part of the sequence of a gene is duplicated.

Dominant and Recessive inheritance

An example of epistasis is the interaction between hair colour and baldness. A gene for total baldness would be epistatic to one for blond hair or red hair. The hair-colour genes are hypostatic to the baldness gene. The baldness phenotype supersedes genes for hair colour, and so the effects are non-additive. (Wikipedia)

• Epistasis: A type of interaction between genes in which the genotype at one locus masks the phenotype or expression of an allele of another gene/ locus.

o Example: A dominant allele for orange coat color (denoted O) can exist on the X chromosome of cats. This dominant orange is separate from the autosomal gene controlling agouti/black coat color (Agouti) and has an epistatic effect on the agouti gene. A male cat (X/Y) with the dominant orange allele (O) will be orange regardless of its genotype at the agouti locus.

• Exon: A section of DNA in a gene that codes for the amino acids of a protein product. After intervening intron sequences have been removed, exons are spliced together to form mature messenger RNA This messenger RNA (or mRNA) is then translated into a protein.

• Expression (of a gene): The phenotypic outcome produced by transcription and translation of a gene; the way in which a gene manifests as a phenotype.

F

• Fixed: An allele is described as being fixed in a population when it is the only allele for a given gene present in the population. In other words, all individuals in the population will be homozygous for that variant.

o Example: One of the defining traits of the Russian Blue cat breed is its dilute gray (“blue”) coat, which is a recessive trait. All Russian Blue cats genotype as d/d, and the dominant D allele is absent from the population. Therefore this breed is fixed for dilute.

• Frameshift mutation: The insertion or deletion of base pairs in a DNA sequence, in which the number of added or deleted base pairs is not a multiple of three, thus shifting the triplet reading frame of the gene product (protein). This results in the synthesis of a protein with amino acids different from those normally specified by the gene

G

• Gene: A sequence of DNA occupying a particular fixed location (locus) on a chromosome. For those genes that code for proteins, the sequence of DNA comprising a gene directs the synthesis of the protein product. Genes are the basic unit of heredity.

• Genome: An organism’s complete DNA sequence found in one set of its chromosomes. This includes the DNA that makes up the 20,000+ genes of mammals as well as all non-coding regions of DNA.

• Genotype: An organism’s set of genes; the genetic makeup of that organism. In practice and in the context of genetic testing, the term “genotype” is

used to reference a particular gene of interest and to specify for the individual being tested what alleles are present at that locus

• Germ line: Cells that pass on genetic information from one generation to the next. Specifically, egg and sperm cells are called germ cells while all other cells of the body are called somatic cells.

• Haplotype: A haplotype is defined as a combination of alleles being inherited together; a set of DNA polymorphisms found on the same chromosome

• Heteroplasmy: Heteroplasmy occurs when there is more than one type of mitochondrial DNA (mtDNA) in an individual.

• Heterozygous: Possessing two different alleles at a particular locus

• Homozygous: Possessing identical alleles at a particular locus. In diploid organisms, which have two copies of each autosomal gene, this means possessing two copies of the same variant

• Incomplete Dominance: A trait is incompletely dominant when the phenotype is dependent on how many copies of the variant allele is present. In other words the phenotype is different if there are zero, one, or two copies of the allele. Typically the heterozygotes have an intermediate phenotype between the two homozygous conditions.

o Example: A horse with a red base color and one copy of the cream allele has an intermediate phenotype of Palomino, while two copies of the cream allele creates a paler Cremello color.

• Incomplete penetrance: Conditions or traits with incomplete penetrance will be expressed in only some of the individuals who possess the trait-associated allele; some individuals with the allele may never express the trait.

• Insertion: The addition of DNA base pairs to an existing DNA sequence. Insertions may be as small as a single base pair or as large as an entire section of chromosome that encompasses multiple genes

• Intron: A section of DNA in a gene that does not code for protein. Introns are intervening sequences between the coding regions of genes, which are called exons. The non-coding introns are spliced out of pre-messenger RNA, leaving only coding exons in the resulting messenger RNA. Each gene may contain multiple introns.

• Intronic mutation: A mutation that occurs in the intron, or portion of a gene that does not code for a protein.

For an excellent explanation of the canine genome please see the website of the NIH (National Human Genome Research Insititute)

https://research.nhgri.nih. gov/dog_genome/canine_ genome.shtml

M

• Lethal mutation (lethal allele): A lethal mutation causes premature death of an organism. Lethal mutations may lead to mortality at any stage of an organism’s development, including after birth, but often act in the early stages of embryonic or fetal development.

o Embryonic lethal: A genotype is considered embryonic lethal if it causes termination of embryonic development. Some individuals with embryonic lethal mutations can survive to term but are nonviable.

• Locus: The location of a particular gene on a chromosome. Plural is loci.

• Mendelian (Mendelian inheritance): Pattern of inheritance that follows the principles proposed by botanist Gregor Mendel. Mechanistically, Mendelian inheritance follows the law of segregation and the law of independent assortment.

o Law of segregation: Alleles segregate when gametes (sperm and egg) are formed during meiosis (division of cells to form specialized reproductive cells). Each gamete thus carries only one version of the gene (one allele).

o Law of independent assortment: The alleles of any one gene will segregate independently from the alleles of other genes (with the exception of genes located close together on a chromosome).

Single locus autosomal dominant, autosomal recessive, and X-linked inheritance patterns follow Mendel’s laws and are said to be Mendelian. Non-Mendelian inheritance patterns can be caused by epistasis and other complex genetic phenomena including multiple genes and environmental components.

• Melanocyte: A specialized cell that produces melanin. Found in the skin, some structures of the eye, the inner ear, and several other tissues.

• Melanoma: A type of cancer, typically in the skin, that arises from changes in an individual’s melanocytes.

• Microsatellite: Regions of repeated DNA found abundantly throughout the genome, often in non-coding areas, and which are highly variable and prone to mutation (see short tandem repeats). The various forms of a given microsatellite (identified by differences in the number of repeats) are referred to as alleles, and these are inherited in a Mendelian fashion. Genetic variation at the microsatellite level can be used to identify individuals (see microsatellite marker).

• Microsatellite marker: Specific microsatellite sequences (short tandem repeats) found at a partic-

ular location on a chromosome that are often used in various kinds of genetic analyses. Microsatellite markers can be used to identify individuals and have applications in forensics, parentage analysis, and studies of populations.

• Missense mutation: A specific type of change in the nucleotide sequence of an organism’s DNA in which a single change in the DNA causes a change in the building block (amino acid) of the gene product (protein).

• Mitochondria: Mitochondria (singular: mitochondrion) are membrane bound organelles, or specialized parts of cells. They play a key role in generating cellular energy known as adenosine triphosphate (ATP). Mitochondria possess their own independent genome separate from the DNA found in the nucleus (see: mitochondrial DNA).

• Mitochondrial DNA: DNA found specifically in the mitochondria. Mitochondrial DNA is a single, circular chromosome. In mammals, this DNA is transmitted primarily from mother to offspring via maternal egg cells.

• Mitotype: Mitochondrial haplotype; a set of variants (alleles) that are inherited together on the mitochondrial DNA

• Mode of inheritance: The manner in which a trait is inherited across generations; the pattern of transmission of a genetic trait.

o Examples: autosomal recessive inheritance, autosomal dominant, X-linked inheritance.

• Modifier: A gene that affects (or modifies) the phenotypic outcome, actions, or product of another gene.

• Mutation: A change in the nucleotide sequence of DNA. A mutation can be small (e.g. single base pair) or large (e.g. segment of a chromosome encompassing multiple genes). Mutations create alleles, which are alternate versions of a gene.

• Nonsense mutation: A specific type of change in the nucleotide sequence of an organism’s DNA in which a single change in the DNA causes a premature stop or truncation of the gene product (protein).

• Nucleotide: An organic molecule that is the basic structural unit of DNA (and RNA). Nucleotides consist of a nitrogenous base, a five-carbon sugar molecule, and a phosphate group. In DNA, the nitrogenous bases are Adenine (A), Guanine (G), Thymine (T), and Cytosine (C). In RNA, Uracil (U) replaces Thymine (T).

• Penetrance: The proportion of individuals in a population that have a particular variant and express the

trait caused by that variant. In other words, when a variant causes a trait but not everyone with the variant has the phenotype, the allele is said to be incompletely penetrant

• Phenotype: The expressed characteristic or trait of an organism; this can include an organism’s appearance, physical features, biochemistry, physiology, and behaviors. An organism’s phenotype can result from the genetic makeup (genotype) or can result from the interaction of genes and the environment.

o Examples of phenotypes: coat color, blood type, disease status, etc.

• Pigment: A compound that selectively absorbs certain wavelengths of light while reflecting others, which creates the appearance of a particular color. In mammals, the pigment eumelanin is responsible for shades of black and brown coloration, and the pigment phaeomelanin is responsible for shades of yellow and red.

• Pleiotropy: Occurs when a single gene affects multiple phenotypic characteristics.

o Example: The mutation responsible for leopard complex spotting in horses affects coat pigmentation as well as vision in low light conditions.

• Polymorphism: The occurrence of two or more different sequence variants at a particular locus. (see MAN diagram)

• Recessive: A trait is recessive when two copies of a variant allele are needed to produce (express) the phenotype

• RNA: Ribonucleic Acid, a single-stranded organic molecule composed of nucleotides. Like DNA, RNA is a nucleic acid made up of a chain of nucleotides; unlike DNA, it is single-stranded instead of double helix, has a ribose sugar backbone instead of a deoxyribose sugar backbone, and contains the nitrogenous base uracil (U) in place of thymine (T). One of the main roles of RNA in the cell is to be the messenger or intermediate of genetic information. DNA is used as the template to make RNA in a process known as transcription, and then RNA is used as a template to make protein products by the process of translation

• Self-color (coat coloration): Term used in cat and dog breeding. The individual hairs of an animal described as “self-colored” are unbanded along the hair shaft, resulting in the appearance of a solid, uniform color from the tip of the hair to the base of the hair. (In contrast, mammals with agouti banded hair have alternating areas of black and red pigments along individual hair shafts, creating a banding pattern.)

• Sex-linked: A gene that is sex-linked resides on one

of the sex-determining chromosomes. In mammals, these are the X or Y chromosomes. Sex-linked traits follow a different inheritance pattern than nonsex-linked traits (autosomal traits) because of the way sex chromsomes are inhertied. In other words males have one X and one Y and females have two X chromosomes. Many sex-linked traits in mammals are X-linked recessive, meaning the causal variant is located on the X chromosome and females need two copies to have the trait where males will only need one.

• Short tandem repeats (STR): A repetitive pattern found in DNA in which a short sequence of nucleotide base pairs, usually 1-6 base pairs, is tandemly repeated as a unit (i.e. one repeat right after the next). STRs occur most frequently in noncoding regions of the genome.

• Single nucleotide polymorphism (SNP): A type of genetic variation in which a single base is substituted at a specific location. For example in the stretch of sequence TAAGTCTGACACAAT if the first G was substituted for a T then the sequence would become TAATTCTGACACAAT. That SNP would be denoted as a G>T variant.

• Splice site: Places in the DNA that have specific information on how a gene is put together. During the process of transcription, DNA is copied into RNA There are regions of the DNA that code for proteins (exons) and non-coding regions that are intervening (introns). At the junction of intron-exon boundaries, there is specific sequence information that tells the cellular machinery to remove (splice) the introns out of the RNA and bring the exons together. Splice sites, then, are where splicing occurs at these intron-exon boundaries. During splicing, the non-coding introns are removed from precursor messenger RNA (pre-mRNA), which then produces mature messenger RNA (mature mRNA) that contains only coding exons. Changes in the genetic sequence of a splice site can affect splicing and subsequently produce an altered gene product.

• Splice variant: A mutation that disrupts the normal splicing, or removal of an intron, that occurs when a gene is transcribed

• Transcription: The molecular process by which a DNA sequence is transcribed into RNA (usually messenger RNA, mRNA).

• Translation: The molecular process by which transcribed RNA directs the synthesis of a protein product by encoding a sequence of amino acids.

• Variant: An alternate form or version of a gene, also known as an allele. A variant is defined as a change in the DNA sequence when comparing individuals in a population.

• Wild type: The wild type version of a gene (the wildtype allele) is the allele that existed before a mutation occurred in the population.

• X-linked: A gene that is X-linked resides on the X chromosome

o Because males (X/Y genotype) have only one X chromosome, they will express an X-linked trait if they inherit one copy of the trait-associated allele, regardless of whether the mode of inheritance is dominant or recessive.

o In contrast, females (X/X genotype) will only express a X-linked recessive trait if they inherit two copies of the trait-associated allele.

• Zygote: The single cell formed by the union of gametes (sperm and egg); a fertilized egg.

Cystinuria

The most common genetically inherited disorder in the Irish Terrier breed today.

“A genetic disorder is a disease caused in whole or in part by a change in the DNA sequence away from the normal sequence. Genetic disorders can be caused by a mutation in one gene (monogenic disorder), by mutations in multiple genes (multifactorial inheritance disorder), by a combination of gene mutations and environmental factors, or by damage to chromosomes (changes in the number or structure of entire chromosomes, the structures that carry genes).”

(Source: National Human Genome Research Institute, webpage Overview)

Cystinuria in Dogs and Cats: What Do We Know after Almost 200 Years?

By Simona Kovaříková, Petr Maršálek, and Kateřina Vrbová

Abstract

Simple Summary

Cystinuria, as an inborn error of metabolism, is a problem with worldwide distribution and has been reported in various canine and feline breeds. Transepithelial transport of cystine is mediated by COLA transporter and the mutation in genes coding this transporter may cause cystinuria. Urolithiasis associated with typical clinical signs may be the clinical consequence of cystinuria. The mutation causing cystinuria and the mode of inheritance have been determined only in several canine breeds. This makes cystinuria difficult to control and gradually decreases its prevalence. In cats, cystinuria occurs only rarely.

The purpose of this review is to summarize current knowledge on canine and feline cystinuria from available scientific reports. Cystinuria is an inherited metabolic defect characterized by abnormal intestinal and renal amino acid transport in which cystine and the dibasic amino acids ornithine, lysine, and arginine are involved (COLA). At a normal urine pH, ornithine, lysine, and arginine are soluble, but cysteine forms a dimer, cystine, which is relatively insoluble, resulting in crystal precipitation. Mutations in genes coding COLA transporter and the mode of inheritance were identified only in some canine breeds. Cystinuric dogs may form uroliths (mostly in lower urinary tract) which are associated with typical clinical symptoms. The prevalence of cystine urolithiasis is much higher in European countries (up to 14% according to the recent reports) when compared to North America (United States and Canada) where it is approximately 1–3%. Cystinuria may be diagnosed by the detection of cystine urolithiasis, cystine crystalluria, assessment of amino aciduria, or using genetic tests. The management of cystinuria is aimed at urolith removal or dissolution which may be reached by dietary changes or medical treatment. In dogs with androgen-dependent cystinuria, castration will help. In cats, cystinuria occurs less frequently in comparison with dogs.

1. Introduction

Cystinuria is an inherited disorder characterized by the impaired reabsorption of cystine in the proximal tubule of the nephron and the gastrointestinal epithelium. The defective transport also involves the

Find this article on Google by searching: Canine Cystinuria

National Library of Medicine

National Ceneter for Biotechnology Information

Pubmed

Paola Scarpa, Academic Editor

Published online 2021 Aug 19

Kovaříková S, Maršálek P, Vrbová K. Cystinuria in Dogs and Cats: What Do We Know after Almost 200 Years? Animals (Basel). 2021 Aug 19;11(8):2437. doi: 10.3390/ani11082437. PMID: 34438894; PMCID: PMC8388795.

Simona Kovaříková

Department of Animal Protection and Welfare and Veterinary Public Health, Faculty of Veterinary Hygiene and Ecology, University of Veterinary Sciences, 612 42 Brno, Czech Republic

Petr Maršálek

Department of Animal Protection and Welfare and Veterinary Public Health, Faculty of Veterinary Hygiene and Ecology, University of Veterinary Sciences, 612 42 Brno, Czech Republic

Kateřina Vrbová

Faculty of Veterinary Medicine, University of Veterinary Sciences, 612 42 Brno, Czech Republic

Epithelial cells are a type of cell that covers the inside and outside of the surfaces of your body. They are found on your skin, blood vessels, and organs, including your urinary tract. “transport across an epithelial cell can be divided into those that do not require a protein, and those that are protein-mediated”

(J.

D. Stockard, Seldin and Giebish’s The Kidney, 2013)

Dimer a molecule or molecular complex consisting of two identical molecules linked together.

Uroliths (also known as calculi or stones) are solidified aggregates of mineral and non- mineral crystalloids that form in the urinary tract.

Proximal tubules are responsible for reabsorbing approximately 65% of filtered load and most, if not all, of filtered amino acids, glucose, solutes, and low molecular weight proteins. Proximal tubules also play a key role in regulating acid-base balance by reabsorbing approximately 80% of filtered bicarbonate. (see diagram below).

other dibasic amino acids ornithine, lysine, and arginine. The term COLA (cystine, ornithine, lysine, arginine) is used for all these amino acids. Nevertheless, only cystinuria results in urolithiasis because these dibasic amino acids are relatively soluble in urine, despite the fact they may reach high concentrations in the urine of affected animals.

In humans, cystinuria was first described by Wollaston in 1810 when he extracted a large cystolith from one of his patients. He named it cystic oxide, because he believed that it had such chemical properties and that the stone had originated from the bladder wall. Although it was later shown not be an oxide nor secreted from the urinary bladder, when isolated, the amino acid was named cystine in recognition of this historical discovery. In 1908, Sir Archibald Garrod suggested cystinuria as a condition that may be an inborn error of metabolism and Dent and Rose hypothesized that cystinuria is an inborn error of cystine transport.

In dogs, the presence of cystine uroliths was reported in 1823 and thus cystinuria was the first reported canine inborn error of metabolism. Now, cystinuria is a disease with worldwide distribution and it is known to affect more than 170 canine breeds according to the reports of veterinary urolith analysis laboratories. Cystinuria is not a problem of dogs and cats only. Indeed, it has also been reported in other canids and felids or ferrets. The purpose of this review is to summarize the current knowledge on cystinuria in dogs and cats.

2. Etiopathogenesis of Cystinuria

The intestinal epithelium is a critical component of the gut barrier. Composed of a single layer of intestinal epithelial cells (IECs) held together by tight junctions, this delicate structure prevents the transfer of harmful microorganisms, antigens, and toxins from the gut lumen into the circulation.

(Williams et al., Epithelial Cell Shedding and Barrier Function, 2015, Sage Vetrinary Pathology)

A protein homodimer is formed by two identical proteins. A protein heterodimer is formed by two different proteins.

Cystine is a non-essential sulfur-containing amino acid composed of two molecules of the amino acid cysteine. Cystine is absorbed through the wall of the small intestine and is normally present in low concentrations in plasma. Plasma amino acids are freely filtered at the glomerulus (see diagram). Under normal conditions, more than 99% of these amino acids are reabsorbed in the proximal renal tubules. The reabsorption of cystine, ornithine, lysine, and arginine is mediated by COLA transporter.

2.1. COLA Transporter

The COLA transporter (b0,+) was originally thought to be a heterodimer, but is likely a heterotetramer formed from two heterodimers consisting of the rBAT (extracellular heavy chain, encoded by SLC3A1, solute carrier family 3 member 1) and b0,+AT (the light chain, encoded by SLC7A9, solute carrier family 7 member 9) subunits joined by a disulfide bridge. This system is the main effector of cystine reabsorption in the kidney. The apical transport system b0,+ mediates influx dibasic amino acids and cystine in exchange

for neutral amino acids. The subunit b0,+AT has 12 transmembrane domains typical cell membrane transporters and heterodimers with rBAT exclusively to form the COLA amino acid transporter. Mutations in either rBAT or b0,+AT can cause cystinuria. In people, 133 mutations in SLC3A1 and 95 mutations in SLC7A9 have been identified. Reported mutations include nonsense, missense, splicing, frameshifts, and large sequence rearrangements.

The same defect is present in the epithelial cells of the small intestine and altered transport of COLA amino acids from gastrointestinal tract occurs, but this is of little consequence as amino acids are primarily absorbed as small peptides. With the exception of lysine, these amino acids are classified as non-essential, and all four of these dibasic amino acids may be absorbed in their dipeptide forms from the gastrointestinal tract. Thus, cystinuria is not associated with protein malnutrition or COLA amino acid deficiency. Nevertheless, it can be associated with disorders of other amino acids. Four English Bulldogs and one Miniature Dachshund with cystinuria were diagnosed with carnitine and taurine deficiency while participating in a clinical trial evaluating dietary management of cystine urolithiasis. In three of these dogs, excessive quantities of carnitine were lost in their urine, but urine excretion of taurine was within the reference range in spite of plasma taurine deficiency. It can be explained by the fact, that cystine is a precursor amino acid and increased renal excretion of cystine may adversely affect taurine synthesis.

2.2. Genetic Aspects

In humans, cystinuria has been classified phenotypically into two types: Type A is caused by defects in SLC3A1 and is inherited in a true autosomal recessive manner, with heterozygotes having a normal urinary excretion of cystine. Type B is caused by SLC7A9 variants and is autosomal dominant with incomplete penetrance, with heterozygotes having a variable degree of cystine hyperexcretion, some within the physiologic range.

In dogs, the first genetic description of cystinuria was conducted with Newfoundlands and autosomal recessive inheritance was suggested. Later, the C-to-T nonsense mutation in exon 2 of the SLC3A1 gene was described. This mutation acts as an early stop codon, precluding the production of rBAT protein and leading to the loss of b0,+. In that report, cystinuric dogs of other breeds were examined (Swedish Lapphund, Dachshund, German short-haired pointer, Irish Setter, Jack Russel Terrier, Corgi) and either heterozygosity at the SLC3A1 locus or a lack of mutations coding region of the SLC3A1 gene were observed. This finding indicates that cystinuria is genetically heterogenous in dogs.

Homotetramer on left and heterotetramer on right

Splice mutation, A genetic alteration in the DNA sequence that occurs at the boundary of an exon and an intron (splice site). (NCI dictionary)

A stop codon is a sequence of three nucleotides (a trinucleotide) in DNA or messenger RNA (mRNA) that signals a halt to protein synthesis in the cell. (National Human Genome Research Institute)

Open reading frames (ORFs) are defined as spans of DNA sequence between start and stop codons. (Science Direct Topics)

Base substitutions are the simplest type of gene-level mutation, and they involve the swapping of one nucleotide for another during DNA replication. (Nature)

Brons et al. (2013) performed a study of mutations associated with cystinuria in various breeds of dogs and a new classification system for canine cystinuria was established according to their results. In cystinuric Labrador Retriever and Labrador mix-breed dogs, the deleted guanine in codon GGC causes a shift of the open reading frame, leading to premature stop codon 41 codons later. This leads to truncation of the rBAT protein to 157 amino acids instead of 784. The early termination probably causes accelerated RNA decay and decreased or no protein synthesis. The mutation was identified in an autosomal recessive form of the disease which is phenotypically and genetically similar to that previously described in Newfoundlands. Only homozygous Labrador Retrievers were cystinuric (both males and females, regardless of neuter status) and developed cystine calculi early in life, albeit more frequently and earlier in males. Labrador Retrievers that were heterozygous for this nonsense mutation showed neither signs of the urinary tract disease nor positive nitroprusside test.

In Australian Cattle dogs with cystinuria, a heterozygous deletion of six bases was found in exon 6 of the SLC3A1 gene. The same homozygous 6 bp deletion was found in one cystinuric mixed breed dog. According to the genetic breed determination, this dog consisted of 1/4 each from Miniature Poodle, Chihuahua, and Shih-tzu with several other breeds consisting of the last quarter, but with no evidence of Australian Cattle dog. All Australian Cattle dogs, males and females, homozygous or heterozygous for this mutation were cystinuric. Homozygous males showed clinical signs (urethral obstruction) earlier in life than heterozygous males. Thus, cystinuria in Australian Cattle dogs is inherited as an autosomal dominant trait.

In Miniature Pinschers, a single base change (missense mutation) in exon 9 of the SLC7A9 gene was detected. It caused the substitution of a large positively charged, hydrophilic arginine for the very small, hydrophobic glycine residue. All cystinuric Miniature Pinschers assessed in this study were found to be heterozygous for this mutation. The exonic sequence of the SLC3A1 gene did not indicate any mutation. These results document the heterogeneity in canine cystinuria.

In dogs, cystinuria had been historically divided into two types: type I referring to a mutation in a SLC3A1 gene with autosomal recessive inheritance and non-type I cystinuria which is associated with milder degree of cystinuria and which is observed in mature intact males of various breeds. The new classification system describes type I cystinuria with autosomal recessive inheritance, type II with autosomal dominant inheritance, and type III for sex-limited inheritance. Involvement of the SLC3A1 gene is indicated by adding A, and simi-

larly B indicates mutations in SLC7A9 (Table 1).

Classification system for cystinuria according to Brons et al. 2013 [26]. Phenotype

Mastiff and related breeds

Breeds

Type III cystinuria (androgen-dependent) occurs in intact males only, occurs later in the life when compared to Newfoundlands, is less severe and associated with variable concentrations of cystine in the urine. This type of cystinuria is reported in Mastiffs, French and English Bulldogs, Basset Hounds, and Irish Terriers. An SLC3A1 nonsense mutation appears to be associated with cystinuria in Mastiffs and related breeds, but not in Irish Terriers or Scottish Deerhounds. The underlying cause of this proposed androgen dependency is currently unknown.

Nevertheless, in most affected breeds, the mutation causing cystinuria is not determined. In several breeds, sequencing of the exons of both SLC3A1 and SLC7A9 did not identify any putative underlying mutation. Thus, further studies of various affected breeds are needed to detect the mutation and to determine the mode of inheritance.

3. Cystine Urolithiasis

3.1. Solubility of Cystine in Urine

Cystinuria by itself does not result in urolithiasis and many cystinuric dogs do not form uroliths. Major factors involved in urolith formation include supersaturation of urine with calculogenic minerals resulting in crystal formation, effects of urinary inhibitors and promoters of crystallization, crystal aggregation, and growth. Cystine uroliths formation is affected mainly by urine pH. Cystine is relatively insoluble at physiological pH levels of 5–7, with a pKa level of 8.3. Up to pH 7, the solubility of cystine is approximately 250 mg/L, whereas at a urine pH level of 7.5, this will double to 500 mg/L urine and triple at pH 8 or higher

3.2. Prevalence of Cystine Urolithiasis

The prevalence of cystine urolithiasis in dogs varies with geographic location and time (details are specified in Table 2).

Table 2

Prevalence of cystine urolithiasis in dogs by region and characteristics of affected dogs in various studies.

For full table please see original file online.

English Bulldog (OR 44.2), Newfoundland (OR 12.6), Dachshund (OR 7.6), Chihuahua (OR 5.6), Miniature Pinscher (OR 9.3), Welsh Corgi (OR 5.0) b

Mastiff (OR 52.7), Australian Cattle Dog (OR 30.8), Pitbull Terrier (OR 12.9), Rottweiler (OR 11.9), English Bulldog (OR 10.1), Bulldog (OR 9.1) c

Females: Pitbull Terrier, crossbreed, Newfoundland

English

Notes from Table 2

a The prevalence of cystine urolithiasis decreased during the period. b OR = odds ratio. Odds ratio was calculated by logistic regression analysis by comparing breed distributions in dogs with cysteine urolithiasis with breed distributions of 2 groups (dogs with other urolith types and dogs examined at the Veterinary Medical Teaching Hospital at the University of California during the same period as the study). c OR = odds ratio. Odds ratio was calculated by logistic regression analysis by comparing breed distributions in dogs with individual urolith type to mixed breed dogs with the same mineral type. d X-ray microanalysis, infrared spectroscopy, scanning electron microscopy. e Significant decrease of cystine urolith prevalence during the study period. f Significant increase of cystine urolith prevalence during the study period. g Uroliths were analyzed in Minnesota Urolith Centre. h OR = odds ratio. Chi-squared tests were performed to assess whether particular breeds were over-represented among the dogs forming cystine uroliths compared with the national insurance company database. i The prevalence of cystine gradually decreased from 27% between the years 1979–1985 to 5.5% in period from 2000 to 2007. j In 26 cases, the sex was not reported. k Sex was reported in 45 cases only. l In 12 cases, the sex was not provided. m OR = odds ratio. Odds ratio was calculated by logistic regression analysis by comparing of the dogs with cystine uroliths to general population of dogs in Hungary according to the Hungarian Microchip Register. n A gradual increase in cystine uroliths was noted (from 12% in 2010 to 30% in 2018). o In 6 cases, the sex was not reported.

According to the most available reports, the prevalence in North America (United States and Canada) is approximately 1–3%. The prevalence in European countries is much higher, ranging from 3% up to 26% in older reports or up to 14% in the most recent studies. Considering the results of recent studies from North America and Europe, the calculated prevalence of cystine urolithiasis is 0.8% in North America and 8.5% in European countries. When evaluating the trends in prevalence of cystine urolithiasis, a gradual increase can be observed in the last decade, both in North America and Europe. The latest published report from Minnesota Urolith Center mentioned the cystine urolith prevalence of 7% (from total 61,160 submissions). Nevertheless, this high number is at least partly affected by submissions from Europe, where the highest portion of cystine uroliths can be seen when compared to other areas.

In all reports on canine cystinuria, males are affected significantly more often than females (98.8% and 1.2%, respectively; these numbers were calculated by taking together available data from scientific reports). Androgen dependency in type III cystinuria may explain the

epidemiological observation that cystine urolithiasis has historically been more common in dogs from European countries than from the USA, where neutering of dogs is more common. In the United States, most (68%) young adult dogs (1–4 years) is castrated and the percentage of castrated dogs gradually increases with age to 81% in adult dogs (4–10 years) and 86% in dogs older than 10 years. In Germany, 43.1% of canine population older than one year is neutered (39% of males and 48% of females). Similar numbers are reported from England, where 44.73% of male dogs are neutered. Cystine urolithiasis typically occurs in young adult and middle-aged dogs, with reported means from 4 to 6 years.

Various breeds are associated with cystinuria. The most mentioned are English Bulldog, Newfoundland, Dachshund, Chihuahua, Staffordshire Bull Terrier, Rottweiler, French Bulldog, and Miniature Pinscher. Nevertheless, reported breeds vary with the geographical location and time of the study and these results may be affected by the breed popularity. Lulich and Ulrich report more than 170 canine breeds where cystine urolithiasis was diagnosed (without specification). The list of breeds particularly mentioned in scientific reports is in Table 3

Table 3

The list of canine breeds where cystinuria was reported

Canine Breeds

Afghan French Bulldog Pug

Akita Inu

German Braque Puli

Alaskan Malamute Golden Retriever Rat Terrier

American Staffordshire Terrier Great Dane Rottweiler

Australian Cattle Dog Husky Rough Collie

Australian Shepherd Dog Chihuahua Saluki

Australian Terrier Irish Terrier Samoyed

Basenji Jack Russel Terrier Scottish Deerhound

Basset Hound Kromfohrländer Scottish Terrier

Bichon Frise Labrador Retriever Setter

Border Collie Landseer Shetland Collie

Borzoi Lhasa Apso Shetland Sheepdog

Boxer Maltese Shih Tzu

Brussels Griffon Mastiff Schnauzer

Bull Mastiff Miniature Pinscher Silky Terrier

Cairn Terrier Miniature Poodle Staffordshire Bull Terrier

Cavalier King Charles Spaniel Miniature Schnauzer

Cocker Spaniel

Dachshund

Munsterlander

Newfoundland

Dalmatian Old English Sheepdog

Staffordshire Terrier

Swedish Lapphund

Tibetian Spaniel

Welsh Corgi

Dobermann Pekingese West Highland White Terrier

Drever Pitbull Terrier Whippet

English Bulldog Pointer

Fox Terrier Poodle

Yorkshire Terrier

Cystine uroliths are rarely (3%) reported from the upper urinary tract. The most common localization of cystine uroliths are urinary bladder and urethra. They may cause urethral obstruction with typical clinical manifestation. This is consistent with the findings in mice or ferrets but in contrast with human medicine, where cystine nephroliths are more common. Occasionally, cystine uroliths may be associated with urinary tract infection. Ling et al. (1986) reported the presence of UTI in almost one third of cases.

4. Diagnosis

4.1. Diagnosis of Cystine Urolithiasis

Imaging methods are the most definitive diagnostic tool for detection of urolithiasis in general. They are used to verify the presence of uroliths and their location, number, size, shape, and density. The radiodensity of cystine stones compared to soft tissue is similar to struvite, less than calcium oxalate and calcium phosphate, and greater than urate. Survey radiographs may be insensitive for detection of small cystine uroliths (less than 1 to 3 mm). Double contrast cystography and/or ultrasonography may be needed. However, ultrasonography involves difficulty in detecting uroliths in the ureters and urethra. Thus, the combination of various methods may be necessary.

Canine cystine uroliths are usually round or ovoid shape with smooth surface. The color may vary, e.g., yellowish brown, medium-light tan, and a range from light yellow to reddish brown are reported (Figure 2). They are commonly multiple, e.g., Méric et al. reported seven as a median number of stones. Their size varies from less than a millimeter to several centimeters in diameter. Most canine cystine uroliths are pure and few contain other minerals, especially ammonium urate and calcium oxalate or struvite. In contrast, Escolar et al. reported the presence of small amounts of calcium apatite in at least 55% of canine cystine uroliths.

If your male Irish Terrier lifts a leg to urinate and does not expel any urine, go to the vet immediately, even a 24 hour wait can compromise the condition of the urethra.

Quantitative research means collecting and analyzing numerical data to describe characteristics, find correlations, or test hypotheses. (Scribbr)

The methods of qualitative data collection most commonly used in health research are document study, observations, semi-structured interviews... (BMC Neurological research and practice)

An estimation of the urolith composition may be done on the basis of their macroscopic appearance, but this may be associated with considerable errors. Quantitative methods (optical crystallography

and infrared spectroscopy) are currently methods of choice. Qualitative analysis showed less than 50% agreement in the case of cystine calculi.

4.2. Diagnosis of Cystinuria

4.2.1. Urinalysis

Cystine crystals are colorless hexagonal plates. Their six sides may or may not be equal and the crystals tend to aggregate and appear layered (Figure 3). Their detection in urine provides strong support for cystinuria because these crystals do not occur in healthy animals. It is noteworthy that cystine crystals are not constantly present in cystinuric dogs.

4.2.2. Assessment of Aminoaciduria

A coloric cyanide-nitroprusside test may be performed. Sodium cyanide reduces cystine to cysteine and the free sulfhydryl groups subsequently react with nitroprusside to form a characteristic purple color. Nevertheless, the test requires dangerous substances. Thus, it is not suitable as an in-house test despite being easy to perform and only selected laboratories offer this test.

Direct measurement of urine cystine concentration is the most precise method allowing quantification. The most used techniques are high-pressure liquid chromatography and automated amino acid analyzers. Not all cystinuric dogs show the same pattern of urinary amino acid loss. Some dogs only lose cystine, whereas others demonstraate increased excretion of cystine, as well as ornithine, lysine, and arginine. The difference (isolated cystinuria vs. urinary excretion of other amino acids) may be caused by the genetic background of the disease, i.e., the specific mutation in particular gene and homozygosity or heterozygosity. Genetic variants may affect the impairment of the transmembrane carrier and thus the extent of aminoaciduria. Because of altered tubular reabsorption of the dibasic amino acids associated with cystinuria, the concentration of ornithine, lysine, and

Dibasic- Relating to a compound that contains two basic monovalent groups or atoms.

arginine should be evaluated together with cystine. The results of COLA amino acids may also support diagnosis in the case cystinuria, because cystine may precipitate and thus cause lower concentrations than were originally present in the urine. The urine concentration of amino acids is expressed as micromoles per gram of creatinine. Dogs with either cystine levels of >200 μmol/g creatinine or COLA values of >700 μmol/g creatinine are considered cystinuric. In cystinuric dogs, the urinary cystine excretion seems to be affected by age. Older dogs over five years were found to have significantly lower cystine levels than younger dogs (five years or younger).

4.2.3. Genetic Tests

In some breeds, genetic tests for cystinuria are available (http:// research.vet.upenn.edu/WSAVA-LabSearch, accessed on 1 May 2021). Such tests offer a method of diagnosing cystinuric animals before they present with clinical signs of cystine urolithiasis and may identify not only clinically affected patients, but also asymptomatic carriers. The results may have an impact on breeding programs.

5. Treatment and Prevention

Cystinuria per se, as an inborn error of metabolism, cannot be successfully treated. The management of cystinuria is aimed at urolith removal or dissolution in case of urolithiasis and/or prevention of urolith formation. After surgical removal, cystine uroliths commonly recur within 6–12 months. Because of the high recurrence rate, prevention is necessary. Without such a strategy, many owners may resort to euthanasia instead of further surgical interventions.

Different therapeutic approaches have been described over the years, such as dietary modification, reduction of urine cystine concentration by induced diuresis, increase of cystine solubility by urine alkalinization and conversion of cystine to a more soluble compound with D-penicillamine or tiopronine According to the current recommendations on the treatment and prevention of uroliths, medical dissolution should be considered before removal. In cases when dissolution cannot be achieved (medications or dissolution foods cannot be administered or tolerated or the urolith cannot be adequately bathed in modified urine), minimally invasive techniques for urolith removal should be preferred.

5.1. Dietary Treatment

Dietary treatment plays a crucial role in the management of cystine stone formation. The dissolution can be achieved by decreasing the concentration of crystallogenic compounds and by increasing cystine solubility.

We do not currently have a specific test for the Irish Terrier. Identifying the genetic marker would allow us to erradicate this condition in our breed, as has been accomplished with Hyperkeratosis.

Urine dilution is an essential step for the prevention and/or dissolution of uroliths regardless of their mineral type. Increased diuresis decreases the concentration of crystal precursors and stimulates more frequent urination, decreasing the time for crystal aggregation. Increasing dietary moisture significantly reduces urinary specific gravity and it is an effective way to enhance diuresis.

5.1.1. Protein

High-protein foods should be avoided in dogs at risk of cystine urolithiasis. Consumption of low protein, moist veterinary food led to a 20–25% reduction in 24-h urine cystine excretion in cystinuric dogs when compared to moist, canine adult maintenance food. Urine cystine excretion can be modulated by dietary protein intake, and more specifically methionine (precursor of cysteine) and cysteine. Feeding a diet containing amounts of these essential amino acids close to their minimum is therefore recommended. Most plant protein sources have smaller amounts of sulfur amino acids than animal proteins. Protein levels in foods for cystinuric dogs should be between 10% and 18% dry matter. Because of possible taurine and carnitine deficiency, cystinuric dogs should be monitored or their diets should be supplemented with carnitine and taurine.

5.1.2. Sodium

Dietary sodium restriction seems to be an important component of the therapeutic strategy in cystinuric people because dietary sodium may enhance cystinuria. Dietary sodium in canine therapeutic diets should be limited to less than 0.3% dry matter.

5.1.3. Urinary pH

As mentioned above, the solubility of cystine in urine is pH dependent. Beneficial effect has been reported in feeding alkalinizing food. Thus, the food that produces a urinary pH range of 7.1–7.7 is recommended for dogs with cystine urolithiasis. Urinary pH values higher than 7.7 should be avoided until it is determined whether they provide a significant risk factor for formation of calcium phosphate uroliths. When a therapeutic diet alone is not able to provide alkaline urine, the administration of alkalinizing agents is recommended. Because of the reports that dietary sodium may enhance cystinuria, potassium citrate should be preferred to sodium bicarbonate. Desired goals of dietary management are urine pH values above 7.5 and urine specific gravity below 1.020.

5.2. Medical Treatment

5.2.1. D-Penicillamine

5.2.2. 2-Merkaptopropionyl-glycine (Tiopronin)

5.2.3. Captopril

5.2.4. Bucillamine

5.3. Castration

Surgical or medical castration can resolve cystinuria in the subset of male dogs with androgen-dependent cystinuria. Castration appears to lower the urinary cystine and COLA concentrations and to prevent cystine calculi formation. The effect of castration in breeds with type of cystinuria seems to have greater effect in comparison with dietary changes . To determine whether neutering reduces cystinuria, measurement of urine cystine concentration before and three months after castration is recommended. If the urine cystine remains elevated at three months, another evaluation should be performed again at six months. Persistently positive results indicate that the dog has a non-androgen-dependent form of the disease. In dogs with androgen-dependent cystinuria, the question may be raised as to whether neutering alone will result in urolith dissolution. In dogs with other types of cystinuria, castration should be recommended as well to prevent further breeding (wanted or accidental) and thus the passing of this condition on to the future generations.

5.4. Future Therapies

5.4.1. L-Cystine Dimethyl Ester and L-Cystine Methyl Ester (L-CDME and L-CME)

5.4.2. Alpha-Lipoic Acid

5.4.3. Selenium

5.4.4. Tolvaptan

Common Canine Health Risks

What are the most common health threats for dogs today? Here I would like to take a look at transmitted diseases and conditions.

In synthesis, the agents can be:

• Bacteria

• Fungi

• Protozoa

• Viruses

The modes of transmission can be:

• Airborne infection

• Droplet infection

• Direct contact

• Indirect contact

• Transmission by other organisms

• Fecal-oral

The ASPCA starts the list of the most prevalent canine health problems with cancer and diabetes. Then we find a familiar evil known to all who wait in vet reception rooms and look at the latest pharmaceutical advertisements to prevent heartworms. If the easily preventable condition of heartworms is third on the list of common health conditions in dogs, then the public is clearly not taking the posters seriously enough!

Who hasn’t returned from a dog show with a case of kennel cough? In fourth place is this pernicious virus that soon spreads to

all the other dogs at home, it is like a bad cold, and with care, will disappear. The next on the list, however, is a word that feels like a sword of ice in the heart of any breederparvovirus. Having carefully cautioned new families to keep the puppy safe at home until the full vaccination cycle is complete, there are times when they just couldn’t wait to bring their bundle of joy round to their friends and family, and perhaps a stop at a store on their way.

The phone call when that dreaded word is first pronounced can so easily be the kiss of death to the little puppy you entrusted to loving hands only a week ago! Fortunately, the two cases Tuscan Red has encountered have been miraculously overcome by the dedicated care of the families and the prowess of their vets.

Rabies, a disease that has been nearly eradicated but is 100% fatal, and Ringworm, not a protozoa, but a fungus that affects the skin and can spread to other animals and humans, complete the list, thank goodness we have not yet met these formidable threats. Tuscan Red has had two cases of slipping patella, and 8 of Cystinuria. No other genetically inherited pathology has yet been identified.

Regular Care of your Irish Terrier

Routine care of a dog involves plenty of hands on, loving interaction. A puppy should begin regular ten minute training sessions from the day they arrive in their new home. Training stimulates their mind and is their special time to enjoy your full attention. Twice a day is good, but as the great Jean Donaldson points out, as rarely twice a week can be effective. Learn to train your dog at a quality training center, or by reading any of the authors listed here. Never train when you are tired or irritable or have pressing issues on your mind. The sessions should only be a positive experience for the dog, any other outcome is counterproductive.

The second form of hands-on upbringing necessary for a puppy is cuddling. While this may be less true of some breeds, the Irish Terrier usually feels fulfilled when there are regular ‘attention sessions’ throughout the day. I recommend cuddling your dog when all four feet are on the ground. From my desk chair I can bend over and caress my dog, give kisses and keep her close to me without her jumping up. In our home, evening TV time is the general sofa cuddle time.

Never discipline a young puppy but do provide plenty of down time in the crate. From about eight months of age, discipline can include a sharp tone of voice, a shocked facial expression etc, but never ever hit the dog with your hand or any other object. As Jean Donaldson makes clear, bad behavior is usually due to bad training or the owner’s forgetfulness in leaving objects withing the dog’s reach or providing regular ‘potty breaks’. When a dog acts up, always consider honestly if you could have avoided it with careful attention.

Your Irish should be brushed once a day and stripped once a month. Show dogs are stripped once or twice a week. The optimal grooming routine is to bathe the dog, strip on that or the following day, then apply topical flea medication about three days after the bath. Always keep nails short enough that you cannot hear ‘tap, tap’ on the floor surface.

Homemade meals are a serious undertaking and should not be improvised. A nutrition professional should give you some guidelines and monitor the diet over the years. Prepared dog food is a good alternative today. An Irish Terrier needs a very light diet to avoid any gastrointestinal

issues, so investing in the most expensive lines of dog food is not the best choice for this breed. In Italy I recommend Monge, Hills, Forza 10, and some colleagues have been happy with Wolf. Any ingredients that are not chicken are best, since the bone meal used to prepare the food is where the residual antibiotics of chicken feed are most concentrated. Food is also about pleasure, so the evening meal I prepare usually includes a strip of fish, some raw beef (frozen for at least two weeks’ time or cooked), an egg yolk, a spoonful of greek yogurt, or in a hurry, a sprinkle of parmeson cheese. I serve the bowls with some hot water added at the last moment to maintain the crunchy texture but to guarantee that they have moisture with the dry food. My dogs usually eat in their crates and rest there after their meal for about 30 minutes, so they never feel under pressure to eat quickly. Growing puppies or pregnant dogs have breakfast at Tuscan Red, and that usually has some Greek yogurt on top. I supplement with Mesoflex during the teething period.

These foods are not recommended for dogs. My only exception to this list is dairy products since I do use ricotta, plain greek yogurt, and kefir.

• Alcohol

• Avocados

• Bacon and other kinds of fatty meat

• Candy and gum

• Chocolate

• Dairy products

• Garlic and onions

• Grapes and raisins

• Raw eggs

• Salty food (pretzels and popcorn with too much salt)

• Sugar

• Xylitol (a class of sweetener known as sugar alcohol)

• Yeast dough

Visits to the vet start the week the puppy arrives in your home. The first check up should establish that the vet is a great place to go, I suggest you bring some little treats that the vet can give your puppy during the visit. Bring the pup in the crate, and do not let it walk on the floor at the clinic, since sick dogs usually visit the vet, this is about the most germ intensive place to be. Never leave the puppy unattended, it may try to jump from the table, and after the vet has checked the puppy place it in the crate again. Expect the vet to look at all parts of the puppy, take the temperature, ask for a sample of feces, weigh and check teeth. Remember that Tuscan Red has a collaborative policy. All of the families who adopt my puppies agree not to spay or neuter until at least three years of age, after we have evaluated together whether to use the dog in the Tuscan Red reproduction program. Moreover, as a general rule, neutering or spaying a dog before 18 months of age is not advantageous since sex hormones are part of the growth and development toward full maturity of the organism. (Please note that neutering a dog is never

a corrective choice, it will not alter the behavior of the dog nearly as much as proper training will. If you are planning to adopt a puppy, plan to take the time, on a regular basis for the first six months, to train the dog you want to live with for the next 12 – 16 years!)

If you have not done any blood testing, then a good time might be at about one year of age. After that, unless there are symptoms of poor health, once every two years is sufficient for the first eight years of life, and after that, an annual blood test is a good idea. There are two elements to a blood test, the blood chemistry values (serum), and the hematology values. Resulting values may vary according to the machines used, but to have a general idea of what to expect I have included these two tables.

Complete blood tests (CBC) are usually accompanied by a urine test. The results show information about potential conditions such as anemia, inflammation, infection, and correct functioning of the immune system and clotting ability, and the optimal functioning of the organs and all aspects of the endocrine system. A CBC reveals detailed information, including:

• Hematocrit (HCT): With this test, we can identify the percentage of red blood cells to detect hydration or anemia.

• Hemoglobin and mean corpuscular hemoglobin concentration (Hb and MCHC): These are pigments of red blood cells that carry oxygen.

• White blood cell count (WBC): With this test, we measure the body’s immune cells. Certain diseases or infections can cause WBC to increase or decrease.

• Granulocytes and lymphocytes/monocytes (GRANS and L/M): These are specific types of white blood cells.

• Eosinophils (EOS): These are a specific type of white blood cells that can indicate health conditions due to allergies or parasites.

• Platelet count: (PLT): This test measures cells that form blood clots.

• Reticulocytes (RETICS): High levels of immature red blood cells can point to regenerative anemia.

• Fibrinogen (FIBR): We can glean important information about blood clotting from this test. High levels can indicate a dog is 30 to 40 days pregnant.

(source for CBC list and tables: website of the Carolina Veterinary Specialists, Winston-Salem)

Meet Stefano Chiesi

Stefano Chiesi is a native Tuscan. From a very early age he has loved animals, especially horses which he learned about visiting his grandfather’s stable. He began working with the Maremmana breed, hoping to become a Buttero, but soon he was overwhelmed by the fascination of Dressage. Having demonstrated aptitude for this pursuit, his family agreed to send him to study with Alessandro Alemani in Germany for three years and later to Australia to work with Mathew Dowsley for 1 year.

Back home he began working with sponsors, training horses and showing professionally as a dressage rider. He is currently in a transitional phase searching for a venue where he can house and train dressage horses as well as hold lessons. Dogs have always been a part of life in the stables, and once he felt established in Italy, he started to look around for the right breed for

him. “What I am looking for in any horse….is the desire to work together, to collaborate. Every horse with or without an illustrious genealogy has many gifts, the trick is to communicate and win him as your friend.“ This quote from an article on Stefano in Cavallo magazine (July 2020) seems to reflect his thoughts on choosing a dog. “I was attracted to the Irish Terrier looking at the whole picture, the dimensions, the conformation, the playful yet collaborative character, just the right tone of vivacity.”

In the fall of 2021, he stopped by for a visit to Tuscan Red where TR Summus (aka Banfi) was growing up. This was a singleton puppy from Geordan Principia and Fireflash Jameson, and he was a smashing pup. Stefano was presented to me by Andrea Vallerini whom I have known for years, the daughter of Alessandro and Cinzia, professional groomers, and breeders of show Wire Fox Terriers (Template Kennel). I

had seen Andrea fall in love with the dog world and earn the respect of many colleagues training and working show rings both in Italy and the USA, so let’s just say that I was ready to like Stefano.

Moreover, as we chatted under the loggia with my dogs all crowding around him, Stefano inspired immediate confidence with his easy calm manner and the evident competence he possessed in handling animals. I never thought I would part with Banfi, but I found myself inviting them to think about him. Before a month had gone by they returned to collect Banfi, and a very exciting life began for the little pup. He was introduced to all the other dogs in the stables, and soon made friends with the horses as well. He attended the carnival festivities at Viareggio and took on the whole world with the equanimity of a solid Irish terrier.

Stefano and Banfi conquered the Italian Jr title at the annual winner show in Milan judged by Carla Molinari, Francesco Cochetti, and Geoff Corish. They are now planning his adult campaign but will conclude his Jr career with Crufts 2023. Best of luck to you both!

Z Males

Z Females

Image Credits

Cover: Moigan Azimi

Insert: background by Johannes

P3 TR Uccelliera by Monika Kripp, Genetic marker; site of Dr. Orawan. TR Vigorello by Marco Mancini, TR Ghiaie della Furba e Enrico Bini self portrait.

P 5 TR Summus in the snow photo Stefano Chiesi, image A. Tureen

P 8 Allele image by Biology Dictionary, Chromosomes from Live Science- Chromosomes: Facts about our genetic storerooms, stop codon from Numerade.

P 9 Deletion from Expii – Deena Hauze, dominant and recessive from The Tech Interactive, Epistasis from Wikipedia, Frameshift from Understanding evolution.

P 11 Incomplete dominance by Science Facts, exon Intron by Britannica kids.

P 12 The laws of segregation and Independent assortment by Genetics Rocks you tube.

P 13 Mitochondral DNA from the National Human Genome Research Institute.

P 14 Genotype phenotype from LabXchange, Polymorphism from STechies, DNA RNA from Expii – Gabi Silzewska.

P 15 Transription from Nagwa.

P 16 From ovulation to Implantation from 123RF, Sperm navigation from Verywell Health.

P 18 Transport from ScienceDirect.com.

P 20 Peptide from Peptide Sciences.

P 26 Qualitatative and Quantitatative from Optimal Workshop’s Blog.

P 30 TR Ombre by Massimo Molinari

P 31-32 Field with mist by Max Mei

P 33 Irish in Rome by Manuela Cola

Images not identified are taken by the author or found on internet without specific reference. Any person with information concering the images used here will please contact Anne Tureen who will make the necessary changes.

P 34 Chocolate by Medical News Today

P 35 Taste testing by Paolo Quagliotto

P 37 TR Tiburzio in a snowy Vineyard by Eugenio Barcelona, Christmas dance by Monika Kripp

P 38 TR Ricciotto by Alessandro Scarpa

Back cover: TR Migliara and TR Incontri by Silvia Costa.

Reccommended

Any book by Jean Donaldson

Founder of the Canadian Academy for Dog training, see also her excellent video series Dog Training 101 by The Teaching Company.

Any book by Karen Pryer

Behavioral biologist from the United States.

Any book by Stanley Coren

Neuropsychologist and professor of Psychology in the United States.