Animal medical genetics: a historical perspective on more than 50 years of research into genetic disorders of animals at Massey University.

Over the last 50 years, there have been major advances in knowledge and technology regarding genetic diseases, and the subsequent ability to control them in a cost-effective manner. This review traces these advances through research into genetic diseases of animals at Massey University (Palmerston North, NZ), and briefly discusses the disorders investigated during that time, with additional detail for disorders of major importance such as bovine α-mannosidosis, ovine ceroid-lipofuscinosis, canine mucopolysaccharidosis IIIA and feline hyperchylomicronaemia. The overall research has made a significant contribution to veterinary medicine, has provided new biological knowledge and advanced our understanding of similar disorders in human patients, including testing various specific therapies prior to human clinical trials.

First case of a dog infected with Aspergillus (Phialosimplex) caninus in Australasia.

Case history: A 2-year-old Rottweiler dog from Perth (WA, Australia) was referred for assessment of a chronic productive cough and weight loss.Clinical findings: Severely enlarged bilateral superficial cervical lymph nodes and severely enlarged abdominal organs were present. The body condition score was poor and there was moderate muscle wasting. Thoracic and abdominal computed tomography images revealed severe diffuse enlargement of thoracic and abdominal lymph nodes, hepatomegaly and diffuse splenomegaly. A diffuse bronchial pattern with severe multifocal saccular bronchiectasis was identified in the lungs.Diagnostic findings: Fungal organisms were seen within macrophages on cytological preparations and on histopathological sections of biopsies of the superficial cervical lymph node. Macrophages contained intracytoplasmic, non-filamentous round-to-ovoid organisms, which varied in size from 5-30 µm in diameter with variable morphology. Budding was not observed, and no hyphae were present. Fungal culture of lymph node tissue resulted in growth of Aspergillus (Phialosimplex) caninus which was confirmed by amplification and sequencing of a segment of the 16S-23S rRNA internal transcribed spacer. Concurrent bacterial bronchitis was diagnosed on culture of broncho-alveolar fluid.Diagnosis: Disseminated aspergillosis caused by Aspergillus caninus.Clinical relevance: This is believed to be the first report of infection caused by A. caninus in a dog in Australasia. The dog was treated with itraconazole for 7 months and was still alive 7 months after the start of treatment.

Regional variations in percutaneous absorption of methimazole: an in vitro study on cat skin.

The use of transdermal gel medications in cats has become popular in veterinary medicine due to the ease of administration compared to oral medication. The research to support systemic absorption of drugs after transdermal gel administration and the preferred skin region to apply these drugs in cats is limited. The aim of this study was to characterize the effect of different skin regions on the percutaneous absorption pharmacokinetics of a commercially available transdermal methimazole after a finite dose was applied to feline skin in vitro. A commercial formulation of methimazole (10 mg) was applied to four skin regions (the inner stratum corneum of the ear, groin, neck, and thorax regions) from six cats. The receptor medium was sampled up to 36 h postapplication, and methimazole concentrations were measured using high-performance liquid chromatography. Methimazole was absorbed more completely across the pinnal skin, compared to the groin, neck, and thorax (P < 0.001), which justifies application to the pinna to maximize efficacy and also to minimize the effects of grooming.

Percutaneous absorption of methimazole: an in vitro study of the absorption pharmacokinetics for two different vehicles.

The use of transdermal medications in cats has become popular in veterinary medicine due to the ease of administration compared to oral medication. However, the research to support systemic absorption of drugs applied to the pinna after transdermal administration in cats is limited. The aim of this study was to characterize the percutaneous absorption pharmacokinetics of methimazole in a lipophilic vehicle compared to methimazole in Pluronic(®) lecithin organogel (PLO) using a finite dose applied to feline ear skin in an in vitro Franz cell model. The two formulations of methimazole (10 mg) were applied to the inner stratum corneum of six pairs of feline ears. The receptor medium was sampled up to 30 h post-administration, and methimazole concentrations were measured using high-performance liquid chromatography (HPLC). Histological examination of all ears was undertaken as small differences in the thickness of ear skin may have contributed to inter-individual differences in methimazole absorption between six cats. Methimazole was absorbed more completely across the pinnal skin when administered in the lipophilic vehicle compared to administration in the PLO gel (P < 0.001).

The prevalence of ocular lesions associated with hypertension in a population of geriatric cats in Auckland, New Zealand.

AIMS: To provide an estimate of the prevalence of ocular lesions associated with hypertension in geriatric cats in Auckland, New Zealand and to evaluate the importance of examination of the ocular fundi of cats over eight years of age. METHODS: A total of 105 cats ≥8 years of age were examined and clinical signs recorded. Blood was collected for the laboratory measurement of the concentrations of blood urea nitrogen (BUN), glucose and creatinine in serum, urine was collected for determination of urine specific gravity (USG), and blood pressure (BP) was measured using high definition oscillometry equipment. A cat was determined to have systemic hypertension with a systolic BP ≥160 mm Hg and a diastolic BP ≥100 mm Hg. Each animal had an ocular fundic examination using a retinal camera to diagnose ocular lesions associated with hypertension, including retinopathies, choroidopathies and optic neuropathies. RESULTS: Blood pressure was successfully recorded in 73 cats. Of these, 37 (51%) had no hypertensive ocular lesions and no underlying disease diagnosed, 24 (33%) had no hypertensive ocular lesions detected, but underlying disease such as chronic kidney disease, hyperthyroidism or diabetes mellitus was diagnosed, and 12 (16%) cats had evidence of hypertensive ocular lesions. Ten of the cats with hypertensive ocular lesions were hypertensive at the time of the first visit and two were normotensive. One additional cat had hypertensive ocular lesions, but it was not possible to obtain consistent BP readings in this animal. Chronic kidney disease was the most commonly diagnosed concurrent disease in cats with hypertensive ocular lesions (n=6). Mean systolic BP for cats with hypertensive ocular lesions (168.0 (SE 6.29) mm Hg) was higher than for those with no ocular lesions (144.7 (SE 3.11) mm Hg) or those with no lesions but with underlying disease (146.0 (SE 4.97) mm Hg) (p=0.001). CONCLUSIONS: Ocular fundic examination of cats over eight years of age allows identification of cats with hypertensive ocular lesions, often before the owner or veterinarian is aware the cat has a problem with its vision. This may result in diagnosis of systemic hypertension allowing early treatment and resolution of lesions. CLINICAL RELEVANCE: The current study demonstrates that ocular lesions resulting from hypertension occur frequently enough in cats in Auckland to support the recommendation for fundic examination in cats over eight years of age as part of the routine physical examination.

Choanal atresia in a cat.

CASE HISTORY: A 5-month-old domestic shorthair kitten with a history of chronic left unilateral nasal discharge was examined. CLINICAL FINDINGS: Endoscopy and computed tomography (CT) demonstrated a complete membranous obstruction of the left nasal choana, confirming congenital unilateral choanal atresia. In addition, congenital hydrocephalus was detected on CT. Nasopharyngoscopy was performed for confirmation and treatment of choanal atresia. DIAGNOSIS: Unilateral membranous choanal atresia associated with congenital hydrocephalus. CLINICAL RELEVANCE: This report describes a rare congenital condition and for the first time, CT imaging provided an accurate diagnosis and allowed planning for the successful treatment of unilateral membranous choanal atresia. In addition, concurrent congenital hydrocephalus was diagnosed.

Permanent monoparesis in a dog after intramedullary injection of iohexol into the lumbar spinal cord.

UNLABELLED: Abstract CASE HISTORY: An 8-year-old, spayed, Doberman Pinscher bitch presented for assessment of acute hindlimb paresis. CLINICAL FINDINGS: During a lumbar myelographic contrast study a small volume of iohexol contrast agent was inadvertently injected into the cord parenchyma. After surgical hemilaminectomy for an intervertebral disc extrusion at L1-2 the dog recovered use of one hindlimb, but had ongoing extensor weakness of the left hindlimb. Left femoral nerve function had not returned after 14 months. DIAGNOSIS: EMG findings 14 months after the incident indicated persistent femoral neuropathy consistent with the intramedullary contrast injection at L3-4. CLINICAL RELEVANCE: Inadvertent deposition of iohexol into spinal cord parenchyma may be rare, but if it occurs it can have long-lasting consequences.

A serological survey of leptospiral antibodies in dogs in New Zealand.

AIM: To investigate the prevalence of titres to four endemic leptospiral serovars in dog sera from the lower half of the North Island, and the South Island of New Zealand submitted to diagnostic laboratories, and to explore the association between the prevalence of seropositive samples to leptospirosis and breed group, age group and sex. METHODS: Serum samples from 655 dogs residing in the central and lower North Island and from the South Island of New Zealand were sourced from the Massey University Veterinary Teaching Hospital and from submissions to New Zealand Veterinary Pathology in 2005. They were screened by the Microscopic Agglutination Test (MAT) against Leptospira interrogans serovars Copenhageni and Pomona and L. borgpetersenii serovars Hardjo and Ballum. Titres greater or equal to 96 were considered positive. Variables investigated for their association with the prevalence of seropositive samples to leptospirosis included serovar, breed, North vs. South Island, age and sex. RESULTS: Positive MAT titres to Leptospira interrogans serovar Copenhageni were found in 10.3 % of dogs (95% CI=8.1-12.9), and were more common than positive titres to other leptospiral serovars. Small breeds did not have a lower prevalence of Copenhageni titres than other breeds. Positive titres to Leptospira borgpetersenii serovar Hardjo were associated with breeds of dogs used as farm working dogs. There was no significant difference in the prevalence of positive leptospiral titres between dogs from the North or South Islands. Dogs greater than 12 years of age were less likely to have positive titres to Leptospira than younger dogs. No association was found between positive titres and sex. CONCLUSIONS: Breeds of dogs used as farm working were at greater risk of exposure to Leptospira borgpetersenii serovar Hardjo. Small breeds did not have a lower risk of seropositivity to Copenhageni than farm working breeds. Further study should be undertaken to confirm the prevalence of positive titres to leptospirosis in farm dogs and dogs resident in the South Island. CLINICAL RELEVANCE: The risk of dogs being exposed to Leptospira interrogans serovar Copenhageni, and requirement for vaccination against serovar Copenhageni, cannot be determined by geographical location or breed group. Vaccination against Leptospira borgpetersenii serovar Hardjo is likely to be beneficial in working dogs.

A retrospective study of gastric dilatation and gastric dilatation and volvulus in working farm dogs in New Zealand.

AIMS: To present findings from a case series of gastric dilatation (GD) or gastric dilatation and volvulus (GDV) in working farm dogs in New Zealand that were examined at veterinary clinics, and to identify possible risk factors for GD or GDV in working farm dogs in New Zealand using a case-control study. METHODS: This retrospective study included a case-series and a case-control study. The case series analysed information from 62 case records of GD or GDV in working farm dogs seen between August 2004 and September 2009 at 13 veterinary clinics throughout New Zealand. Cases were classified as GD or GDV if the diagnosis was confirmed by radiography, surgery or post-mortem examination. Details of history and treatment, as well as outcomes, were obtained for each case. For the case-control study, records of 41 working farm dogs with GD or GDV (cases) seen between April 2008 and April 2009, and 82 working farm dogs examined because of trauma over the same period and in the same 13 clinics (controls), were used to model the risk factors for GD or GDV. RESULTS: From the case-series study, 40/62 (65%) cases of GD or GDV that were examined and treated at the veterinary clinics returned to work. Of the 41 dogs where the gastric contents were recorded, 25 (61%) had predominantly food or bones in the stomach, and 26/27 dogs had a history of having eaten meat, bones or scavenged a carcass. The case-control study showed that the significant risk factors for GD or GDV, compared with control dogs presenting with trauma, were breed, age and season. The odds that a case of GD or GDV was a Huntaway, after adjusting for age and season, was 19 times higher than the odds a control was a Huntaway. Gender and bodyweight were not identified as risk factors. CONCLUSIONS AND CLINICAL RELEVANCE: A high proportion of farm working dogs with GD or GDV were successfully treated by veterinarians. The risk of a case of GD or GDV being a Huntaway was significantly higher than for a dog presenting as a trauma case. However the influences of the season of the year, climatic factors and nutritional factors on the pathogenesis need to be identified before adequate preventative measures can be recommended.

Congenital hypothyroidism of dogs and cats: a review.

Congenital hypothyroidism is a rare and underdiagnosed congenital endocrine disorder in dogs and cats and the true incidence is unknown. The disorder may cause a range of clinical signs depending on the primary defect, which affect production of thyroid hormones; some cases present when adult. Hallmark clinical signs of congenital hypothyroidism are mental impairment and skeletal developmental abnormalities, resulting in disproportionate dwarfism; goitre may or may not be present. Documented causes of congenital hypothyroidism in dogs include deficiency of, or unresponsiveness to, thyrotropin-releasing hormone (TRH) or thyroid-stimulating hormone (TSH), thyroid dysgenesis, dyshormonogenesis and iodine deficiency. In cats, TSH unresponsiveness, thyroid dysgenesis, dyshormonogenesis and iodine deficiency have been confirmed. Adequate replacement therapy results in a successful outcome in the majority of cases, especially when started early in life, as permanent developmental abnormalities can be prevented. This review describes reported cases in dogs and cats, diagnostic investigation, and recommendations for treatment.

Genetic diversity among Campylobacter jejuni isolates from pets in Ireland.

Campylobacter jejuni isolates obtained from pets housed in shelters and in private households were subtyped by fla typing (using DdeI and HinfI restriction enzymes) and pulsed-field gel electrophoresis (PFGE) techniques. Composite fla cluster analysis on 78 C jejuni isolates was more discriminative than either single fla typing technique with 39.7 per cent single isolate patterns. PFGE on 52 C jejuni isolates revealed 53.8 per cent single isolate patterns and was the most discriminative method applied. A database of C jejuni subtyping profiles from pets in Ireland was assembled. The presence of genetic heterogeneity detected in the C jejuni subtypes suggests that pets can acquire the organisms from multiple potential sources. In addition, heterogeneity was detected in the C jejuni subtypes obtained by different culture methods within the same pet. There was a link between isolates from dogs in close contact in the same environment, confirming that this is a potential route of infection, and clusters were detected containing both cat and dog C jejuni isolates, suggesting possible interspecies transmission.

The experimental induction of leukoencephalomyelopathy in cats.

Leukoencephalomyelopathy of undetermined etiology has been described in specific pathogen-free cats. A study was established to assess if the long-term feeding of a gamma-irradiated diet could induce this disease. Cats fed exclusively on diet irradiated at 25.7-38.1 kGy ("typical" dose) and 38.1-53.6 kGy (high-end dose), respectively, developed typical lesions with attendant, progressively severe ataxia between study days 140 and 174. The onset of ataxia at day 140 and the number of animals affected at this time were similar in animals fed each ration. A maximum ataxia "score" was first reached by an animal on the high-end dose diet on day 167 and by 2 cats fed the "typical-end" dose diet 21 days later. Ataxic cats and 1 animal euthanized on day 93 prior to the onset of ataxia exhibited varying degrees of Wallerian degeneration in the spinal cord and brain, similar to the spontaneous disease. The elevated total antioxidant status of spinal cord segments and hepatic superoxide dismutase concentration of cats fed typical and high-end treated diets suggested free-radical involvement in the pathogenesis. The significantly elevated peroxide concentrations of the irradiated diets (1,040% and 6,440% of untreated values) may have resulted in increased oxidative insult, a factor possibly exacerbated by the treated diets' reduced vitamin A content. This study has reproduced leukoencephalomyelopathy in cats similar to spontaneous outbreaks by feeding a gamma-irradiated dry diet with elevated peroxide and reduced vitamin A concentrations.

A comparison of different culture methods for the recovery of Campylobacter species from pets.

Five culture methods for the recovery of Campylobacter species (spp.) were evaluated on 361 rectal swabs collected from cats and dogs in Ireland. Speciation using PCR methods was performed on all isolates to assess the sensitivity of each culture method for isolation of Campylobacter spp., and to establish the prevalence of Campylobacter jejuni, C. coli, C. upsaliensis, C. lari and C. helveticus. Overall 163 of 361 (45.2%) samples were confirmed Campylobacter spp. positive. Direct plating onto modified charcoal cefoperazone deoxycholate agar (mCCDA) with cefoperazone, amphotericin and teicoplanin (CAT) selective supplement yielded a significantly higher prevalence of Campylobacter spp. (33.0%) than each of the other four methods (P < or = 0.05). This method was also the most sensitive method for isolation of C. upsaliensis compared with any of the other four methods used in the current study (P < or = 0.05). A direct plating method onto mCCDA agar with CCDA selective supplement and a filtration method onto blood agar after pre-enrichment in CAT supplemented broth yielded lower Campylobacter spp. prevalences of 19.7% and 17.5% respectively. A filtration method onto CAT agar and pre-enrichment in Preston broth before plating onto mCCDA agar were less sensitive for the isolation of Campylobacter spp. Speciation results of Campylobacter isolates revealed the majority of Campylobacter isolates were C. upsaliensis (50.0%) and C. jejuni (41.9%). A small number of isolates were C. coli (2.6%), C. lari (1.5%) and C. helveticus (1.1%). The overall detection of Campylobacter spp. in the 361 pets sampled was significantly increased by using a combination of isolation methods (P < or = 0.05), producing a more accurate determination of the prevalence of Campylobacter spp. in pets in Ireland and of the actual Campylobacter species. As the majority of Campylobacter spp. were recovered by direct plating onto mCCDA agar with CAT supplement, this method is the method of choice if only a single method is selected for isolation of the most common Campylobacter spp. detected in pets and humans.

Prevalence of thermophilic Campylobacter species in household cats and dogs in Ireland.

Rectal swabs were collected from 147 household dogs and 35 household cats, including healthy animals, animals with gastrointestinal signs and animals with a variety of medical and surgical conditions. A combination of selective culture methods was used to optimise the recovery of Campylobacter species, and a PCR was used to confirm their isolation and to identify the species. The overall prevalence of Campylobacter species was 42.9 per cent in the cats and 41.5 per cent in the dogs. Campylobacter upsaliensis was the species most commonly isolated from the dogs and cats, and Campylobacter jejuni was the second most commonly isolated. Particularly high prevalences were detected in the few cats and dogs with diarrhoea, and in the cats and dogs that were six months old or younger.

Characteristics of 234 dog bite incidents in Ireland during 2004 and 2005.

Information was obtained by telephone interview from 100 dog owners whose dog had bitten a person, and from 134 victims of bites by a dog not owned by the victim. Three-quarters of the victims were female and aged from 21 to 60 years. The majority of the dogs were owned, male, two to six years old, over 10 kg in bodyweight and belonged to the popular breeds: collies, cocker/springer spaniels, terrier breeds, Jack Russell terriers, German shepherd dogs, golden retrievers and crossbreeds. The numbers of bites by the different breeds indicated that those that inflicted the most bites were the popular breeds rather than the breeds with any greater propensity to bite. Most attacks were rapid single bites and in 50 per cent of the cases, neither the owner nor the victim was able to identify any signal of the dog's intention to bite. Overall, 21 per cent of the incidents were rated as 'serious' and 2 per cent as 'life threatening'. One fifth of the dogs were euthanased as a result of the incident. Half the incidents required professional medical assistance for the victim. Almost half the incidents took place while the victim was walking or passing close to the dog's territory, or while the victim was interacting with the dog at home.

Leukoencephalomyelopathy in specific pathogen-free cats.

Investigations were carried out on 8 specific pathogen-free cats (5 male and 3 female) from a colony experiencing "outbreaks" of progressive hind limb ataxia in 190 of 540 at-risk animals ranging from 3 months to 3 years old. These studies identified moderate to severe bilateral axonal degeneration within white matter regions of the cervical, thoracic, and lumbar spinal cord and in the white matter of the cerebral internal capsule and peduncle, in the roof of the fourth ventricle and inferior cerebellar peduncle, and in the external arcuate and pyramidal fibres of the medulla. There were varying degrees of accompanying microgliosis, astrocytosis, and capillary hyperplasia. Such a clinicopathologic syndrome, termed feline leukoencephalomyelopathy, has previously been described in cat colonies in Britain and New Zealand, although its etiology has not been determined. The degenerative nature of the lesions and their bilateral distribution suggest possible nutritional, metabolic, or toxic causes. Although these findings provide circumstantial evidence that the exclusive feeding of a gamma-irradiated diet of reduced vitamin A content is associated with the development of the neuronal lesions, further tissue micronutrient and antioxidant analysis will be required to support this hypothesis.

Lethal outbreak of disease associated with feline calicivirus infection in cats.

Recently, in the USA, virulent mutants of feline calicivirus (FCV) have been identified as the cause of a severe and acute virulent systemic disease, characterised by jaundice, oedema and high mortality in groups of cats. This severe manifestation of FCV disease has so far only been reported in the USA. However, in 2003, an outbreak of disease affected a household of four adult cats and an adult cat from a neighbouring household in the UK. Three of the adult cats in the household and the neighbouring cat developed clinical signs including pyrexia (39.5 to 40.5 degrees C), lameness, voice loss, inappetence and jaundice. One cat was euthanased in extremis, two died and one recovered. A postmortem examination of one of the cats revealed focal cellulitis around the right hock and right elbow joints. The principal finding of histopathological examinations of selected organs from two of the cats was disseminated hepatocellular necrosis with mild inflammatory infiltration. Immunohistology identified FCV antigen in parenchymal and Kupffer cells in the liver of both animals and in alveolar macrophages of one of them. In addition, calicivirus-like particles were observed by electron microscopy within the hepatocytes of one cat. FCV was isolated from two of the dead cats and from the two surviving cats. Sequence analysis showed that they were all infected with the same strain of virus, but that it was different from strains of FCV associated with the virulent systemic disease in cats in the USA. The outbreak was successfully controlled by quarantine in the owner's house.

Characterization of certain inflammatory variables in the peripheral blood of clinically healthy dogs.

Many laboratory techniques have been developed to study and quantify the inflammatory response, including the release of acid hydrolase enzymes, leukotriene B(4) (LTB(4)) production, reactive oxygen species (ROS) production and complement conversion studies. Although extensively studied in human health and disease, the relevance of such tests in the dog is largely unknown. After isolation of the peripheral blood mononuclear cell (PBMC) and polymorphonuclear cell (PMN) fractions from the peripheral blood of 38 clinically healthy dogs, values for ROS production were similar for both cell fractions when measured by luminol-enhanced chemiluminescence (17,853+/-9,695 U/10(6) cells versus 19,138+/-14,569 U/10(6) cells for the PBMC (n=38) and PMN (n=18) fractions, respectively). However, the mean time taken to reach maximum chemiluminescence was noticeably shorter in the PBMC fraction (5.1+/-3.3 versus 10.7+/-2.5 min for PBMCs (n=36) and PMNs (n=18), respectively). Intracellular concentrations of beta-glucuronidase, beta-galactosidase and N-acetyl-beta-glucosaminidase were assayed by spectrofluorometry. Mean values for all three enzymes were higher in PBMCs (n=31-35) than in PMNs (n=10-14). Both cell fractions released 20% of the intracellular enzyme concentration when stimulated with opsonized zymosan. Following incubation with A23187 (1 microM), mean LTB(4) production was higher in PBMCs (4.45+/-2.92 ng/10(6) cells; n=27) than in PMNs (0.96+/-2.22 ng/10(6) cells; n=13) using a validated high performance liquid chromatography (HPLC) assay. Immunoprecipitation studies revealed that the mean percentage conversion of C3 to C3b following stimulation with opsonized zymosan was 57.3+/-13.4% (n=36). The results provide normal values for clinically healthy dogs that may subsequently be used in future studies investigating dogs with various inflammatory disorders.