Genomic analysis of canine pneumoviruses and canine respiratory coronavirus from New Zealand.

AIMS: To isolate canine respiratory coronavirus (CRCoV) and canine pneumovirus (CnPnV) in cell culture and to compare partial genomic sequences of CRCoV and CnPnV from New Zealand with those from other countries. METHODS: Oropharyngeal swab samples from dogs affected by canine infectious respiratory disease syndrome that were positive for CnPnV (n = 15) or CRCoV (n = 1) by virus-specific reverse transcriptase quantitative PCR (RT-qPCR) in a previous study comprised the starting material. Virus isolation was performed in HRT-18 cells for CRCoV and RAW 264.7 and Vero cells for CnPnV. The entire sequence of CnPnV G protein (1,266 nucleotides) and most (8,063/9,707 nucleotides) of the 3' region of CRCoV that codes for 10 structural and accessory proteins were amplified and sequenced. The sequences were analysed and compared with other sequences available in GenBank using standard molecular tools including phylogenetic analysis. RESULTS: Virus isolation was unsuccessful for both CRCoV and CnPnV. Pneumovirus G protein was amplified from 3/15 (20%) samples that were positive for CnPnV RNA by RT-qPCR. Two of these (NZ-048 and NZ-049) were 100% identical to each other, and 90.9% identical to the third one (NZ-007). Based on phylogenetic analysis of the G protein gene, CnPnV NZ-048 and NZ-049 clustered with sequences from the USA, Thailand and Italy in group A, and CnPnV NZ-007 clustered with sequences from the USA in group B. The characteristics of the predicted genes (length, position) and their putative protein products (size, predicted structure, presence of N- and O-glycosylation sites) of the New Zealand CRCoV sequence were consistent with those reported previously, except for the region located between open reading frame (ORF)3 (coding for S protein) and ORF6 (coding for E protein). The New Zealand virus was predicted to encode 5.9 kDa, 27 kDa and 12.7 kDa proteins, which differed from the putative coding capacity of this region reported for CRCoV from other countries. CONCLUSIONS: This report represents the first characterisation of partial genomic sequences of CRCoV and CnPnV from New Zealand. Our results suggest that the population of CnPnV circulating in New Zealand is not homogeneous, and that the viruses from two clades described overseas are also present here. Limited conclusions can be made based on only one CRCoV sequence, but the putative differences in the coding capacity of New Zealand CRCoV support the previously reported variability of this region. The reasons for such variability and its biological implications need to be further elucidated.

Disseminated Rasamsonia argillacea infection in a dog.

CASE HISTORY: A 4-year-old, male neutered Borzoi presented for unlocalised pain and frequent episodes of vocalisation. CLINICAL FINDINGS: Pain was localised to the lumbar spine and radiographs revealed a L3-L4 lesion consistent with discospondylitis. The dog was treated for presumptive bacterial discospondylitis with surgical debridement, spinal stabilisation, and cephalexin. Samples collected from the affected intervertebral disc at the time of surgery revealed lymphoplasmacytic inflammation with no causative agent identified on histopathology or bacterial culture. After an initial period of improvement, signs recurred despite an 8-week antibiotic course, with the development of inappetence, weight loss, polydipsia, and polyuria. Repeat radiographs revealed a new cervical intervertebral lesion, and concurrent pyelonephritis was diagnosed based on blood and urine results. Fungal culture of urine resulted in growth of Rasamsonia argillacea species complex and disseminated fungal disease was clinically diagnosed. Antifungal treatment was commenced, however the dog deteriorated, and euthanasia was performed. PATHOLOGICAL FINDINGS: Multifocal white plaques were grossly visualised in the spleen, mesenteric lymph nodes, cervical vertebrae, and kidneys. Periodic acid-Schiff-positive, fine, parallel-walled, occasionally branching, septate hyphae 5-10 µm in diameter, and conidia 5-7 µm in diameter were found on sectioning all organs. R. argillacea species complex was identified by fungal culture of urine and was considered the species of fungal organism seen histologically. The isolate was subsequently confirmed as R. argillacea by DNA sequencing. DIAGNOSIS: Disseminated Rasamsonia argillacea infection. CLINICAL RELEVANCE: Rasamsonia argillacea species complex is a recognised invasive mycosis in veterinary medicine, with disseminated disease causing significant clinical complications and death. This is believed to be the first report of infection caused by R. argillacea in a dog in Australasia and highlights the importance of awareness of a potential fungal aetiology in dogs with discospondylitis.Abbreviations: CLSI: Clinical and Laboratory Standards Institute; CRI: Constant rate infusion; MEC: Minimum effective concentration; MIC: Minimum inhibitory concentration; PAS: Periodic acid-Schiff.

A molecular survey of canine respiratory viruses in New Zealand.

AIMS: The aim of this study was to identify viruses associated with canine infectious respiratory disease syndrome (CIRDS) among a population of New Zealand dogs. METHODS: Convenience samples of oropharyngeal swabs were collected from 116 dogs, including 56 CIRDS-affected and 60 healthy dogs from various locations in New Zealand between March 2014 and February 2016. Pooled samples from CIRDS-affected (n = 50) and from healthy (n = 50) dogs were tested for the presence of canine respiratory viruses using next generation sequencing (NGS). Individual samples (n = 116) were then tested by quantitative PCR (qPCR) and reverse transcriptase qPCR (RT-qPCR) for specific viruses. Groups were compared using Fisher's exact or χ2 tests. The effect of explanatory variables (age, sex, type of household, presence of viral infection) on the response variable (CIRDS-affected or not) was tested using RR. RESULTS: Canine pneumovirus (CnPnV), canine respiratory coronavirus (CRCoV), canine herpesvirus-1 (CHV-1), canine picornavirus and influenza C virus sequences were identified by NGS in the pooled sample from CIRDS-affected but not healthy dogs. At least one virus was detected by qPCR/RT-qPCR in 20/56 (36%) samples from CIRDS dogs and in 23/60 (38%) samples from healthy dogs (p = 0.84). CIRDS-affected dogs were most commonly positive for CnPnV (14/56, 25%) followed by canine adenovirus-2 (CAdV-2, 5/56, 9%), canine parainfluenza virus (CpiV) and CHV-1 (2/56, 4% each), and CRCoV (1/56, 2%). Only CnPnV (17/60, 28%) and CAdV-2 (14/60, 23%) were identified in samples from healthy dogs, and CAdV-2 was more likely to be detected healthy than diseased dogs (RR 0.38; 95% CI = 0.15-0.99; p = 0.045). CONCLUSIONS: The frequency of detection of viruses traditionally linked to CIRDS (CAdV-2 and CPiV) among diseased dogs was low. This suggests that other pathogens are likely to have contributed to development of CIRDS among sampled dogs. Our data represent the first detection of CnPnV in New Zealand, but the role of this virus in CIRDS remains unclear. On-going monitoring of canine respiratory pathogens by NGS would be beneficial, as it allows rapid detection of novel viruses that may be introduced to the New Zealand canine population in the future. Such monitoring could be done using pooled samples to minimise costs. CLINICAL RELEVANCE: Testing for novel respiratory viruses such as CnPnV and CRCoV should be considered in all routine laboratory investigations of CIRDS cases, particularly in dogs vaccinated with currently available kennel cough vaccines.

Lack of protection against feline immunodeficiency virus infection among domestic cats in New Zealand vaccinated with the Fel-O-Vax® FIV vaccine.

Infections with feline immunodeficiency virus (FIV) are common in New Zealand, although the impact of those infections on the health status of the cats remains unclear. Although many cats are vaccinated yearly with a commercial FIV vaccine containing FIV subtypes A and D, the effectiveness of this vaccine in protection against infection with field FIVs is unclear, as a high proportion of New Zealand viruses belong to subtype C. The objective of the study was to compare the frequency of FIV infection among adult FIV-vaccinated and FIV-unvaccinated domestic cats with access to outdoors. Buccal swabs were collected by the participating veterinarians and tested for the presence of FIV provirus by quantitative PCR. Overall, 26/185 (14.0 %) samples were positive for FIV, including 7/82 (8.5 %) samples from FIV-unvaccinated and 19/103 (18.4 %) from FIV-vaccinated cats. There was no protective effect of vaccination on FIV infection among sampled cats (p = 0.05). Partial sequences of the FIV envelope gene from five New Zealand viruses were analysed by the maximum likelihood method. All clustered with other New Zealand FIV sequences from subtypes A (n = 2), C (n = 2) or putative recombinant viruses (n = 1). While the FIV vaccination did not prevent FIV infection among sampled cats, it may have had an impact on transmissibility of the virus or on disease progression. As neither was addressed in the current study, further research is needed to fully assess the potential benefits of FIV vaccination. Considering the frequency of FIV infection in FIV-vaccinated cats, FIV infection status should be monitored not only before the first vaccination, but before each yearly booster.

A serological survey of canine respiratory coronavirus in New Zealand.

Aims: To determine the seroprevalence of canine respiratory coronavirus (CRCoV) in New Zealand dogs, and to explore associations with age, sex, breed, month, and geographical region of sampling and reported presence of clinical signs suggestive of respiratory disease.Methods: A total of 1,015 canine serum samples were randomly selected from submissions to a diagnostic laboratory between March and December 2014, and were analysed for CRCoV antibodies using a competitive ELISA. Logistic regression analysis was used to determine associations between seroprevalence of CRCoV and breed category, age, sex, sampling month, region, and reported health status of dogs.Results: Overall, 538/1,015 (53.0%) samples were seropositive for CRCoV, with 492/921 (53.4%) positive dogs in the North Island and 46/94 (49%) in the South Island. Age of dog, sampling month, region, and presence of abnormal respiratory signs were included in the initial logistic regression model. Seroprevalence was higher in dogs aged ≥3 compared with ≤2 years (p < 0.01). The lowest seroprevalence was observed in July (30/105; 28.5%) and August (32/100; 32%), and the highest in June (74/100; 74%). Seroprevalence in dogs from Auckland was higher than in dogs from the Hawkes Bay, Manawatu, Marlborough, and Waikato regions (p < 0.05). Abnormal respiratory signs (coughing, nasal discharge, or sneezing) were reported for 28/1,015 (2.8%) dogs sampled. Seroprevalence for CRCoV tended to be higher among dogs with respiratory signs (67.9 (95% CI = 47.6-83.4)%) than dogs with no reported respiratory signs (52.6 (95% CI = 49.5-55.7)%).Conclusions: Serological evidence of infection with CRCoV was present in more than half of the dogs tested from throughout New Zealand. Differences in CRCoV seroprevalence between regions and lack of seasonal pattern indicate that factors other than external temperatures may be important in the epidemiology of CRCoV in New Zealand.Clinical relevance: Our data suggest that CRCoV should be included in investigations of cases of infectious canine tracheobronchitis, particularly if these occur among dogs vaccinated with current vaccines, which do not include CRCoV antigens.

Untargeted metabolic profiling of dogs with a suspected toxic mitochondrial myopathy using liquid chromatography-mass spectrometry.

'Go Slow myopathy' (GSM) is a suspected toxic myopathy in dogs that primarily occurs in the North Island of New Zealand, and affected dogs usually have a history of consuming meat, offal or bones from wild pigs (including previously frozen and/or cooked meat). Previous epidemiological and pathological studies on GSM have demonstrated that changes in mitochondrial structure and function are most likely caused by an environmental toxin that dogs are exposed to through the ingestion of wild pig. The disease has clinical, histological and biochemical similarities to poisoning in people and animals from the plant Ageratina altissima (white snakeroot). Aqueous and lipid extracts were prepared from liver samples of 24 clinically normal dogs and 15 dogs with GSM for untargeted liquid chromatography-mass spectrometry. Group-wise comparisons of mass spectral data revealed 38 features that were significantly different (FDR<0.05) between normal dogs and those with GSM in aqueous extracts, and 316 significantly different features in lipid extracts. No definitive cause of the myopathy was identified, but alkaloids derived from several plant species were among the possible identities of features that were more abundant in liver samples from affected dogs compared to normal dogs. Mass spectral data also revealed that dogs with GSM have reduced hepatic phospholipid and sphingolipid concentrations relative to normal dogs. In addition, affected dogs had changes in the abundance of kynurenic acid, various dicarboxylic acids and N-acetylated branch chain amino acids, suggestive of mitochondrial dysfunction.

A veterinary perspective on preventing injuries and other problems that shorten the life of working dogs.

Domesticated dogs are widely used for farm work, law enforcement, military service and in assistance roles. Farm dogs assist farmers by guarding or moving stock, reducing the requirement for human workers and enabling steep terrain to be productive. Working and service dogs are subject to illness and injury, depending on their breed characteristics, activities and the environment in which they work. There are limited data in the veterinary literature on the causes of loss and early retirement of working dogs, and targeted research is required to better determine factors that affect their incidence. Many breeds of working dog suffer from inherited forms of musculoskeletal disease; most importantly, elbow and hip dysplasia. The effectiveness of traditional radiographic (phenotypic) scoring schemes at reducing the prevalence of hip and elbow dysplasia has been variable. The selection of breeding stock should preferably be based on estimated breeding values, rather than on an individual phenotypic score/grade. Nutrition can affect physical and mental performance, disease risk and susceptibility, and recovery from injury, age-associated decline and longevity. The dietary proportions of fat, protein and carbohydrate influence fuel selection during exercise. A common concern among farmers is the difficulty of maintaining body condition in their dogs during periods of heavy work with current feeding practices. Additionally, maintaining a positive calcium balance is necessary for adaptive remodelling in response to exercise. Research strategies should include improved treatment options, the development of more specific means of genetic selection for inherited diseases, investigation into specific nutritional requirements and the dissemination of this knowledge.

Les chiens domestiqués sont très largement utilisés pour travailler dans les fermes, ainsi qu'en tant que chiens policiers, chiens militaires ou chiens d'assistance. Les chiens de ferme aident les éleveurs à garder ou à déplacer leurs troupeaux, ce qui réduit les besoins en main d'oeuvre et permet d'exploiter des terrains escarpés. Les chiens de travail et aidants sont plus ou moins exposés à la maladie et aux blessures, en fonction des caractéristiques de leur race, des tâches qui leur sont confiées et de l'environnement dans lequel ils travaillent. Il n'existe pas beaucoup d'études vétérinaires publiées sur les causes des morts et des mises à la retraite prématurées des chiens de travail, thème qu'il conviendrait d'explorer plus précisément afin de mieux déterminer les facteurs affectant l'incidence de ces problèmes. Plusieurs races canines sont atteintes de troubles musculo-squelettiques héréditaires, dont le plus important est la dysplasie du coude et de la hanche. Les méthodes traditionnelles d'interprétation radiologique (mesure des phénotypes) ont contribué à réduire la prévalence de la dysplasie du coude et de la hanche, avec des résultats variables. La sélection des reproducteurs devrait reposer sur l'estimation de leur potentiel génétique (valeur d'élevage) plutôt que sur le score ou l'échelle de mesure du phénotype d'un individu. L'alimentation joue un rôle sur les performances physiques et mentales, la sensibilité aux maladies et le risque sanitaire, les capacités de rétablissement après une blessure, le processus de vieillissement et la longévité. La proportion relative des lipides, protéines et glucides dans l'alimentation donnée aux chiens détermine les sources d'énergie utilisées durant l'effort. Une préoccupation courante parmi les éleveurs concerne la difficulté de maintenir leurs chiens en bonne condition physique pendant les périodes où le travail est le plus dur, compte tenu des pratiques d'alimentation actuelles. Il est également important de maintenir un bon équilibre en calcium afin de soutenir les capacités d'adaptation à l'effort et de récupération. Les stratégies de recherche devraient explorer les pistes d'amélioration des traitements, mettre au point des méthodes de sélection génétique axées spécifiquement sur les maladies héréditaires et mieux élucider les besoins nutritionnels des chiens, sans oublier de diffuser par la suite les connaissances ainsi obtenues.

El uso del perro doméstico en tareas agrícolas, policiales, militares y de socorro o ayuda está muy extendido. Los perros pastores ayudan al granjero vigilando o conduciendo el ganado, lo que reduce la necesidad de personal humano y permite aprovechar las tierras situadas en terrenos escarpados. Los perros de trabajo y de servicio están sujetos a enfermedades y lesiones que dependen de las características de la raza, la actividad de que se trate y el medio en que esta discurra. En la bibliografía veterinaria hay escasa información sobre las causas que subyacen a la pérdida o la jubilación anticipada de perros de trabajo. De ahí la necesidad de investigar específicamente este aspecto para determinar con más precisión los factores que influyen en ello. Muchas razas de perros de trabajo padecen formas hereditarias de enfermedad musculoesquelética, la más importante de ellas la displasia de codo o cadera. Los tradicionales sistemas de puntuación radiológica (fenotípica) han resultado de eficacia variable para reducir la prevalencia de estas dos displasias. La selección de ejemplares reproductores debería basarse no tanto en la puntuación o grado fenotípico de cada individuo como en los valores estimados de cría. La alimentación puede influir en el rendimiento físico y mental, el riesgo de enfermedades y la sensibilidad a ellas, la recuperación tras una lesión y la longevidad o la decadencia ligada a la edad. Las proporciones de grasas, proteínas y carbohidratos presentes en el régimen alimentario influyen en la selección de combustible metabólico durante la actividad física. Uno de los aspectos que preocupa a todos los granjeros es la dificultad de mantener el cuerpo del perro en condiciones en los periodos de mucho trabajo con las actuales prácticas de alimentación. Además, también es necesario mantener un saldo positivo de calcio para que tenga lugar la remodelación adaptativa del cuerpo en respuesta al ejercicio físico. Los planes de investigación deben incluir aspectos como la búsqueda de mejores alternativas de tratamiento, la obtención de medios más específicos de selección genética respecto de las enfermedades hereditarias, el estudio de necesidades nutricionales específicas y la difusión de cuanto conocimiento resulte de estas investigaciones.

Creation of a predictive equation to estimate fat-free mass and the ratio of fat-free mass to skeletal size using morphometry in lean working farm dogs.

AIMS To develop an equation that accurately estimates fat-free mass (FFM) and the ratio of FFM to skeletal size or mass, using morphometric measurements in lean working farm dogs, and to examine the association between FFM derived from body condition score (BCS) and FFM measured using isotope dilution. METHODS Thirteen Huntaway and seven Heading working dogs from sheep and beef farms in the Waikato region of New Zealand were recruited based on BCS (BCS < 3, 3-4, > 4) using a nine-point scale. Bodyweight, BCS, and morphometric measurements (head length and circumference, body length, thoracic girth, and fore and hind limb length) were recorded for each dog, and body composition was measured using an isotopic dilution technique. A new variable using morphometric measurements, termed skeletal size, was created using principal component analysis. Models for predicting FFM, leanST (FFM minus skeletal mass) and ratios of FFM and leanST to skeletal size or mass were generated using multiple linear regression analysis. RESULTS Mean FFM of the 20 dogs, measured by isotope dilution, was 22.1 (SD 4.4) kg and the percentage FFM of bodyweight was 87.0 (SD 5.0)%. Median BCS was 3.0 (min 1, max 6). Bodyweight, breed, age and skeletal size or mass were associated with measured FFM (p<0.001). There was a good correlation between predicted FFM and measured FFM (R2=0.96), and for the ratio of predicted FFM to skeletal size and measured values (R2=0.99). Correlation coefficients were higher for the ratio FFM and leanST to skeletal size than for ratios using skeletal mass. There was a positive correlation between BCS-derived fat mass as a percentage of bodyweight and fat mass percentage determined using isotope dilution (R2=0.65). CONCLUSIONS AND CLINICAL RELEVANCE As expected, the predictive equation was accurate in estimating FFM when tested on the same group of dogs used to develop the equation. The significance of breed, independent of skeletal size, in predicting FFM indicates that individual breed formulae may be required. Future studies that apply these equations on a greater population of working Huntaway and Heading dogs are needed to establish the utility of these equations on a large scale. Such studies could ascertain if there is a ratio for lean mass to skeletal size below which the risk of injury or disease increases. If these equations prove useful they would provide an objective and non-invasive measure to determine when welfare in individual dogs is compromised by underfeeding.

A survey of canine respiratory pathogens in New Zealand dogs.

AIMS To determine which of the common canine respiratory pathogens circulate among selected populations of healthy and diseased dogs in New Zealand. METHODS Coagulated blood samples for serology and oropharyngeal swabs for virology were collected from healthy dogs (n=47) and from dogs with acute respiratory disease (n=49). For diseased dogs a convalescent blood sample was also collected 3-4 weeks later. Oropharyngeal swabs were subjected to virus isolation and tested for canine parainfluenza virus (CPIV), canine adenovirus (CAdV) 2, canine herpesvirus (CHV), canine respiratory coronavirus (CRCoV), canine influenza virus (CIV), canine distemper virus (CDV), Bordetella bronchiseptica, Streptococcus equi subsp. zooepidemicus, and Mycoplasma cynos nucleic acids by quantitative PCR (qPCR). Sera were tested for CRCoV antibody using competitive ELISA and results expressed as percent of inhibition (POI). RESULTS The mean age of diseased dogs (2.7, min <0.5, max 8.5 years) was lower than the mean age of healthy dogs (5.3, min <0.5, max 17 years) (p<0.001). In total, 20/94 (21%) dogs were positive for at least one agent by qPCR. Diseased dogs were most commonly positive for M. cynos (8/47, 17%), followed by CPIV (3/47, 6%) and B. bronchiseptica (3/47, 6%), while healthy dogs were most commonly positive for CAdV-2 (6/47, 13%), followed by M. cynos (2/47, 4%). All samples were negative for CIV, CRCoV, CDV and S. equi subsp. zooepidemicus. Viruses were not isolated from any of the samples tested. In total, 47/93 (50%) dogs were seropositive for CRCoV on at least one sampling occasion. Samples from diseased dogs were more frequently seropositive for CRCoV, with higher POI, than samples from healthy dogs. CONCLUSIONS AND CLINICAL RELEVANCE We showed that CAdV-2, CPIV, CHV, CRCoV, B. bronchiseptica and M. cynos circulated among sampled dogs. The convenience sampling methodology, with a poor match between the populations of diseased and healthy dogs in terms of age, breed and use, together with the relatively small sample size precluded inference of any causal relationships between infection with a given pathogen and development of disease. None-the-less, our data suggest that further investigation into epidemiology and disease association of CRCoV and M. cynos is warranted. In addition, circulation of novel respiratory pathogens among dogs in New Zealand should be considered in future studies, as 70/94 (74%) diseased dogs were negative for all the pathogens tested.

An epidemiological investigation of an idiopathic myopathy in hunting dogs in New Zealand.

AIM: To conduct an epidemiological investigation of an idiopathic myopathy, known as "Go Slow" (GSM), which was initially recognised in dogs used for pig hunting. A secondary aim was to describe the hunting activities, diet and health of dogs used for pig hunting in New Zealand. METHODS: A retrospective cohort study was conducted between June 2014-June 2017. Cases of GSM in dogs were diagnosed by veterinarians using a combination of clinical history, physical examination findings, serum biochemistry and/or skeletal muscle histology. A telephone interview was conducted with the owner or primary veterinarian to provide information regarding the dog's diet and exercise over the 7 days preceding the onset of clinical signs. In August 2015, a separate online survey of owners of dogs used for pig hunting was conducted to characterise the normal hunting activities, diet and health of these dogs. RESULTS: A total of 86 cases of GSM were recruited, of which 58 (67%) were pig hunting dogs, 16 (19%) pet dogs and 12 (14%) working farm dogs. Cases were most commonly reported in the upper North Island, and 65 (76 (95% CI=67-85)%) were from the Northland region. Processed commercial dog food had been fed to 93 (95% CI=88-98)% of affected dogs. Ingestion of raw, frozen or cooked wild pig in the preceding week was reported for 76 (88 (95% CI=82-95)%) dogs with the myopathy. In the survey of owners of healthy pig hunting dogs, 203 eligible responses were received; pig hunting was reported to most commonly occur in Northland (20.2%), Waikato (22.3%) and Bay of Plenty (23.2%) regions. Commercial dog food was fed to 172 (85 (95% CI=80-90)%) of the dogs included in this survey, and 55 (27 (95% CI=20-33)%) had eaten wild pig in the preceding week. The most common reported health problem in pig hunting dogs was traumatic wounds. CONCLUSIONS: Cases of GSM were most commonly recognised in dogs used for pig hunting, but also occurred in pet and working farm dogs. The disease was most frequently reported in the upper North Island of New Zealand and ingestion of wild pig was a consistent feature in cases of this myopathy. CLINICAL RELEVANCE: To minimise the risk of dogs developing this myopathy, it would seem prudent to avoid feeding any tissues from wild pigs to dogs in areas where the disease is known to occur.

Macronutrient intake of dogs, self-selecting diets varying in composition offered ad libitum.

The diet of the domestic dog has changed significantly from that of its wolf ancestor, with to date only two studies having examined macronutrient self-selection in dogs. Whilst the first focused solely on protein intake, determining an intake of 30% metabolisable energy (ME), the second investigated dietary protein, fat and carbohydrate (PFC), indicating an intake ratio of 30:63:7% by energy. This study's aim was to further elucidate macronutrient intake by providing greater macronutrient range, energy content, and to investigate over a longer duration than previous studies. Fifteen adult dogs were given access to three wet diets providing 500% of daily ME, twice daily over 10 days. The diets were nutritionally complete and formulated using the same four ingredients in different proportions to supply high levels of protein (58% ME), fat (86% ME) or carbohydrate (54% ME). Overall fat and carbohydrate consumption significantly declined from 6,382 to 917 kcals per day (p < 0.001) and 553 to 214 kcals day-1 (p < .01) respectively. Protein intake, however, remained constant over the study and ranged from 4,786 to 4,156 kcals day-1 . Such results impacted on percentage total energy intake, with fat decreasing from 68% to 52% (p < .001) and protein increasing from 29% to 44% (p < .01). Our findings suggest that dogs still possess a "feast or famine" mentality, wherein energy dense fat is prioritised over protein initially. With continued feeding over 10 days, a transition to a more balanced energy contribution from both macronutrients is evident. The study also shows that given the option, dogs do not select carbohydrate to be a significant portion of the diet. The health implications of such dietary selection are of interest.

Biological Variability in Serum Cortisol Concentration Post-adrenocorticotropic Hormone Stimulation in Healthy Dogs.

BACKGROUND: The ACTH stimulation has low sensitivity for the diagnosis of hypercortisolism possibly as a result of biological and analytical variability. HYPOTHESIS/OBJECTIVES: To report the components of biological and analytical variability in serum cortisol concentration post-ACTH stimulation ([cortisol]) in healthy dogs. ANIMALS: Fourteen healthy harrier hound dogs. METHODS: The data were extracted from a separate, prospective, randomized, double-blinded, controlled discovery study in which dogs treated with vehicle control and 4 different doses of cortisone acetate (CA) for 7 days had an ACTH stimulation test performed to confirm the dose-dependent effect of CA. The index of individuality (IoI), the critical difference between sequential measurements (CD ), and the number of measurements required to assess the homeostatic set point (HSP) of [cortisol] with confidence intervals (CI) of 90 and 95% were estimated. RESULTS: The IoI was equal to 1.1 and the CD was 3.3 µg/dL (92 nmol/L). The number of measurements required to assess the HSP of [cortisol] with CI of 90 and 95% were 3 and 15, respectively. Additionally, mean [cortisol] was higher in males than in females (13.3 ± 4 µg/dL [366 ± 114 nmol/L] vs. 11.5 ± 2.5 µg/dL [318 ± 65 nmol/L], respectively; P = .046). As expected, treatment with CA resulted in a dose-dependent suppression of [cortisol]. CONCLUSIONS AND CLINICAL IMPORTANCE: False-negative test results in hypercortisolism could occur when [cortisol] is outside of the individual's HSP and within the reference interval. The large CD emphasizes the importance of assessing clinically relevant parameters in the diagnosis and monitoring of HC.

Subclinical Bacteriuria in Older Cats and its Association with Survival.

BACKGROUND: Bacterial urinary tract infections are uncommon in cats in general but the prevalence increases to 29% in older cats with comorbidities (Veterinary Clinical Pathology 2008, 37, 317; Journal of Feline Medicine & Surgery 2007, 9, 124; Veterinary Microbiology 2009, 136, 130). Frequently, the infections are subclinical. The clinical relevance of subclinical bacteriuria (SB) is uncertain, and the optimal treatment requires clarification. OBJECTIVE: Prospective, observational study to: (i) identify the prevalence and incidence count of SB in older (≥7 years), nonazotemic cats, (ii) evaluate specific risk factors for SB, and (iii) investigate the potential relationship between untreated SB and survival. ANIMALS: Sixty-seven, nonazotemic cats were tested on 5 occasions over 3 years. METHODS: Urine samples were obtained by cystocentesis for quantitative urine culture and blood samples for measurement of serum creatinine concentration. Episodes of SB were not treated. Serum creatinine concentration, body weight, urine specific gravity, sex, and age were evaluated as potential risk factors for a positive urine culture. The association between urine culture results and survival was evaluated with Cox's proportional hazard model. RESULTS: A total of 256 urine samples was obtained. The prevalence of SB varied between 10 and 13%, and incident infections were uncommon. Female cats were 21 times more likely to have a positive urine culture than were male cats (odds ratio [OR], 21.2; confidence interval [CI], 4.1-110; P = .00028). Subclinical bacteriuria was not significantly associated with survival. CONCLUSION AND CLINICAL IMPORTANCE: Subclinical bacteriuria is common in nonazotemic, older cats. Although antimicrobial treatment was withheld, the presence of SB was not adversely associated with survival.

Policies for the vaccination of cats and dogs in New Zealand veterinary practices.

AIMS: To determine current practices and attitudes towards vaccination of dogs and cats of veterinarians in New Zealand; the methods used for informing clients on which vaccines to use, and the preferred site for vaccination of cats. METHODS: A postal questionnaire was sent to all 483 listed veterinary practices in New Zealand during February 2012. Some questions were specific to pet dogs, cats, or working farm dogs. Responses were categorised according to practice type and geographical region of the respondent. Factors associated with respondent recommendation of annual vaccination with modified live viral (MLV) vaccines were examined using logistic regression analysis. Vaccines that were considered to be essential for every animal were defined as core; those that may be recommended for animals whose location or lifestyle placed them at risk, were defined as non-core. RESULTS: There were 204 useable returns, equivalent to a response rate of 42.2%, distributed across the country. Annual vaccination with MLV vaccines of dogs was recommended by 54/198 (27.3%) respondents, and of cats by 107/181 (59.1%) respondents. Factors associated with the recommendation of annual administration of MLV vaccines to dogs included being a companion animal practice, a desire for policies on vaccination to be left to individual clinics, and having one veterinarian in the practice. Administration of the final vaccination for puppies was recommended at ≥14 weeks old by 55/185 (29.7%) respondents, and for kittens at ≥13 weeks old by 42/183 (23%) respondents. Of respondents that administered MLV vaccines annually, 62/103 (60.2%) believed reducing the frequency of vaccination would reduce income, and 52/103 (50.5%) considered it would have a negative effect on animal health. Advice to enable clients to decide which non-core vaccines were administered was given by 181/199 (91%) respondents. Factors considered when recommending a vaccine included consideration of risk to individual patients (190/203; 93.6%), requirements of boarding kennels/catteries (165/203; 81.3%) and clinic vaccination policy (142/203; 70%). The preferred site for administering MLV vaccines to cats was the dorsal neck or inter-scapular region (137/198; 69.2%). Amongst respondents, 18 wanted disease surveillance information to allow for truly informed decisions to be made about vaccination. CONCLUSIONS AND CLINICAL RELEVANCE: Veterinarians can now compare their own vaccination practices and attitudes with those of veterinarians nationally, and internationally. There is a need for national surveillance information and for continued education of the public and commercial kennel and cattery owners for optimal vaccination strategies to be developed.

Canine parvoviruses in New Zealand form a monophyletic group distinct from the viruses circulating in other parts of the world.

Canine parvovirus 2 (CPV-2) is a well-recognized cause of acute haemorrhagic enteritis in dogs worldwide. The aim of the current study was to identify which CPV-2 subtypes circulate among dogs in New Zealand, and to investigate the evolutionary patterns of contemporary CPV-2 viruses. Faecal samples were collected from 79 dogs with suspected CPV-2 infection over the period of 13 months, and tested for the presence of CPV-2 DNA by PCR. Of 70 positive samples, 69 were subtyped as CPV-2a and one as CPV-2. A majority of CPV-2 positive samples were collected from unvaccinated or not-fully vaccinated puppies ≤6 months of age. The haplotype network produced from New Zealand CPV-2 sequences showed no structure when assessed based on location, vaccination status or age of the animals sampled. International haplotype network indicated that, unlike CPV-2 from other countries, the population of CPV-2 in New Zealand appeared to be monophyletic.

Quantitative accuracy of the simplified strong ion equation to predict serum pH in dogs.

BACKGROUND: Electrochemical approach to the assessment of acid-base states should provide a better mechanistic explanation of the metabolic component than methods that consider only pH and carbon dioxide. HYPOTHESIS/OBJECTIVES: Simplified strong ion equation (SSIE), using published dog-specific values, would predict the measured serum pH of diseased dogs. ANIMALS: Ten dogs, hospitalized for various reasons. METHODS: Prospective study of a convenience sample of a consecutive series of dogs admitted to the Massey University Veterinary Teaching Hospital (MUVTH), from which serum biochemistry and blood gas analyses were performed at the same time. Serum pH was calculated (Hcal+) using the SSIE, and published values for the concentration and dissociation constant for the nonvolatile weak acids (Atot and Ka ), and subsequently Hcal+ was compared with the dog's actual pH (Hmeasured+). To determine the source of discordance between Hcal+ and Hmeasured+, the calculations were repeated using a series of substituted values for Atot and Ka . RESULTS: The Hcal+ did not approximate the Hmeasured+ for any dog (P = 0.499, r(2) = 0.068), and was consistently more basic. Substituted values Atot and Ka did not significantly improve the accuracy (r(2) = 0.169 to <0.001). Substituting the effective SID (Atot-[HCO3-]) produced a strong association between Hcal+ and Hmeasured+ (r(2) = 0.977). CONCLUSIONS AND CLINICAL IMPORTANCE: Using the simplified strong ion equation and the published values for Atot and Ka does not appear to provide a quantitative explanation for the acid-base status of dogs. Efficacy of substituting the effective SID in the simplified strong ion equation suggests the error lies in calculating the SID.

The serological response of working farm dogs to a vaccine containing Leptospira interrogans serovars Copenhageni and Pomona, and L. borgpetersenii serovar Hardjo.

AIMS: To evaluate the serological response in dogs to a commercial vaccine for use in cattle containing cultured strains of Leptospira interrogans serovars Copenhageni and Pomona, and L. borgpetersenii serovar Hardjo. METHODS: Blood samples were obtained from 67 working farm dogs on 12 farms, and the microscopic agglutination test (MAT) was used to measure titres to the Leptospira spp. serovars Pomona, Hardjo, and Copenhageni. Samples with a titre of <1:25 were defined as seronegative. Dogs that were seronegative to both Pomona and Hardjo (n=33) were randomised to either the vaccination (n=20) or control (n=13) groups. Seven of these dogs were seropositive to Copenhageni. Vaccinated dogs were given the three-component vaccine subcutaneously on two occasions, 4 weeks apart. MAT titres were measured again in both groups 2 weeks after the second vaccination. RESULTS: Of the vaccinated dogs, 20/20 (100%) developed titres to serovar Pomona, and 16/20 (80%) had titres >100; for serovar Hardjo, 19/20 (95%) dogs had titres, with 18/20 (90%) being >100; and for serovar Copenhageni, 15/17 (88%) dogs that were initially seronegative had titres, with 6/17 (35%) being >100. The median titres for Pomona and Hardjo (200 (95% CI=179-359) and 200 (95% CI=176-379), respectively) were lower than for Copenhageni (50 (95% CI=26-124)) in dogs originally seronegative (p<0.001). There was no association between titres to the different serovars. Of the 13 unvaccinated dogs, two developed titres to serovar Pomona, and three to Hardjo. All titres were <100. CONCLUSIONS: The tested vaccine was effective in raising antibodies to the three serovars, although the titres do not guarantee protection. CLINICAL RELEVANCE: There has been a recent increase in cases of leptospirosis attributed to the serovar Pomona in dogs in New Zealand, but the vaccines licensed for use in dogs in New Zealand for the prevention of leptospirosis only protect against Copenhageni/Icterohaemorrhagiae. The vaccine tested in this study produced titres to Pomona and Hardjo that suggest the vaccine could be useful for reducing disease caused by these serovars in working dogs in New Zealand.

Sequence variation of the feline immunodeficiency virus genome and its clinical relevance.

The ongoing evolution of feline immunodeficiency virus (FIV) has resulted in the existence of a diverse continuum of viruses. FIV isolates differ with regards to their mutation and replication rates, plasma viral loads, cell tropism and the ability to induce apoptosis. Clinical disease in FIV-infected cats is also inconsistent. Genomic sequence variation of FIV is likely to be responsible for some of the variation in viral behaviour. The specific genetic sequences that influence these key viral properties remain to be determined. With knowledge of the specific key determinants of pathogenicity, there is the potential for veterinarians in the future to apply this information for prognostic purposes. Genomic sequence variation of FIV also presents an obstacle to effective vaccine development. Most challenge studies demonstrate acceptable efficacy of a dual-subtype FIV vaccine (Fel-O-Vax FIV) against FIV infection under experimental settings; however, vaccine efficacy in the field still remains to be proven. It is important that we discover the key determinants of immunity induced by this vaccine; such data would compliment vaccine field efficacy studies and provide the basis to make informed recommendations on its use.

Isoflavone metabolism in domestic cats (Felis catus): comparison of plasma metabolites detected after ingestion of two different dietary forms of genistein and daidzein.

Some felid diets contain isoflavones but the metabolic capacity of cats toward isoflavones is relatively unknown, despite the understanding that isoflavones have divergent biological potential according to their metabolite end products. The objective of this study was to determine the plasma metabolites detectable in domestic cats after exposure to 2 different dietary forms of isoflavones, either as a soy extract tablet (n = 6) or as part of a dietary matrix (n = 4). Serial blood samples were collected after isoflavone exposure to identify the plasma metabolites of each cat. Genistein was detected in its unconjugated form or as a monosulfate. Daidzein was detected as both a mono- and disulfate as well as in its unconjugated form. Other daidzein metabolites detected included equol mono- and disulfate, dihydrodaidzein, and O-desmethylangolensin. No ß-glucuronide metabolites of either isoflavone were detected. Equol was produced in markedly fewer cats after ingestion of a soy extract tablet as a single oral bolus compared with cats consuming an isoflavone-containing diet. The detectable metabolites of the isoflavones, genistein and daidzein, in domestic cat plasma after dietary ingestion has been described in the present study for the first time. The metabolic capacity for isoflavones by domestic cats appears to be efficient, with only minimal proportions of the ingested amount detected in their unconjugated forms. This has implications for the potential of isoflavones to exert physiological activity in the domestic cat when consumed at concentrations representative of typical dietary intake.

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.