Clinical enzymology of the dog and cat

Clinical enzymology studies the enzyme activity in serum or other body fluids for the diagnosis, prognosis or monitoring of a variety of diseases. Clinical enzymology has greatly benefited from advances in technology and is now an integral part of laboratory analysis. However, to maximise the clinical benefits of serum enzyme measurement, clinicians and clinical pathologists must have a good understanding of the pathophysiology behind serum enzyme alterations. They must also be aware of the preanalytical and analytical factors that can affect the accuracy of serum enzyme activity measurement. This review article first covers the basic concepts of clinical enzymology and the general mechanisms related to serum enzyme alterations. Then, the review discusses the potential effects of various preanalytical and analytical factors on enzyme activity measurement. Lastly, it explores the pathophysiology and clinical use of various serum enzymes in canine and feline medicine. The present review article aims to be a comprehensive one-stop source for clinical pathologists and small animal practitioners.</p

Hypoalbuminaemia and its association with disease and clinical outcomes in cats

Objectives: To report the incidence of feline hypoalbuminaemia and characterise the distribution of presenting disease categories and pathoaetiologies of hypoalbuminaemia in cats. The secondary aim was to evaluate the relationship between hypoalbuminaemia and clinical outcomes. Materials and Methods: Medical records of cats with hypoalbuminaemia (&lt;28.0 g/L, reference interval: 28.0 to 39.0 g/L) presenting to a veterinary teaching hospital over 5 years were retrospectively reviewed. The severity of hypoalbuminaemia was further stratified into mild (24.0 to 27.9 g/L), moderate (20.0 to 23.9 g/L) and severe (≤19.9 g/L) groups. The median albumin and severity groups were compared between the determined disease categories, pathoaetiologies and clinical outcomes. Results: The incidence of hypoalbuminaemia was 32.7% (533/1632). Gastrointestinal disease was the most common disease category associated with hypoalbuminaemia [154/533 (28.9%)], of which, 49.4% (76/154) of cats had gastrointestinal neoplasia. Neoplastic [159/533 (29.8%)] and inflammatory conditions [158/533 (29.6%)] were common pathoaetiologies noted. Statistically significant differences in the serum albumin between individual disease and pathoaetiological categories were found. Cats with moderate to severe hypoalbuminaemia had a statistically significantly longer hospitalisation period, cost of treatment and increased odds of death (odds ratio 2.4, 95% confidence interval: 1.3 to 4.6 and odds ratio 3.2, 95% confidence interval: 1.5 to 6.6, respectively). Clinical Significance: The incidence of feline hypoalbuminaemia in our study surpasses previous canine reports. Our findings support albumin as a negative acute phase protein in cats, with hypoalbuminaemia frequently associated with inflammatory disease. Hypoalbuminaemia also features prominently in cats with gastrointestinal neoplasia, indicating careful appraisal of the presence of protein-losing enteropathy is required in these cases. Finally, albumin is found to be a prognostic indicator in this study.</p

Increased serotonin transporter expression reduces fear and recruitment of parvalbumin interneurons of the amygdala

Genetic association studies suggest that variations in the 5-hydroxytryptamine (5-HT; serotonin) transporter (5-HTT) gene are associated with susceptibility to psychiatric disorders such as anxiety or posttraumatic stress disorder. Individuals carrying high 5-HTT-expressing gene variants display low amygdala reactivity to fearful stimuli. Mice overexpressing the 5-HTT (5-HTTOE), an animal model of this human variation, show impaired fear, together with reduced fear-evoked theta oscillations in the basolateral amygdala (BLA). However, it is unclear how variation in 5-HTT gene expression impacts on the microcircuitry of the BLA to change behavior. We addressed this issue by investigating the activity of parvalbumin (PV)-expressing interneurons (PVINs), the biggest IN population in the basal amygdala (BA). We found that increased 5-HTT expression impairs the recruitment of PVINs (measured by their c-Fos immunoreactivity) during fear. Ex vivo patch-clamp recordings demonstrated that the depolarizing effect of 5-HT on PVINs was mediated by 5-HT2A receptor. In 5-HTTOE mice, 5-HT-evoked depolarization of PVINs and synaptic inhibition of principal cells, which provide the major output of the BA, were impaired. This deficit was because of reduced 5-HT2A function and not because of increased 5-HT uptake. Collectively, these findings provide novel cellular mechanisms that are likely to contribute to differences in emotional behaviors linked with genetic variations of the 5-HTT

Converging Prefronto-Insula-Amygdala Pathways in Negative Emotion Regulation in Marmoset Monkeys

BACKGROUND: Impaired regulation of emotional responses to potential threat is a core feature of affective disorders. However, while the subcortical circuitry responsible for processing and expression of fear has been well characterized, the top-down control of this circuitry is less well understood. Our recent studies demonstrated that heightened emotionality, as measured both physiologically and behaviorally, during conditioned fear and innate/ social threat was induced, independently, by excitotoxic lesions of either the anterior orbitofrontal cortex (antOFC) or ventrolateral prefrontal cortex (vlPFC). An important outstanding question is whether the antOFC and vlPFC act on common or distinct downstream targets to regulate negative emotion. METHODS: The question was addressed by combining localized excitotoxic lesions in the PFC of a nonhuman primate and functional neuroimaging ([$^{18}$F]fluorodeoxyglucose positron emission tomography) with a fear-regulating extinction paradigm. Marmoset monkeys with unilateral lesions of either the antOFC or vlPFC were scanned immediately following exposure to a fearful or safe context, and differences in [$^{18}$F]fluorodeoxyglucose uptake were evaluated. RESULTS: [$^{18}$F]fluorodeoxyglucose uptake in the insula and amygdala of the intact hemisphere was significantly increased in response to the fearful context compared with the safe context. Such discrimination between the two contexts was not reflected in the activity of the insula-amygdala of the antOFC or vlPFC-lesioned hemisphere. Instead, uptake was at an intermediate level in both contexts. CONCLUSIONS: These findings demonstrate that the distinct control functions of the antOFC and vlPFC converge on the same downstream targets to promote emotion regulation, taking us closer to a mechanistic understanding of different forms of anxiety.This research was supported by a Medical Research Programme Grant (No. MR/M023990/1) from the Medical Research Council (to ACR) and was carried out within the Behavioural and Clinical Neuroscience Institute supported by a consortium award from the Wellcome Trust and the Medical Research Council

Brainstem phaeohyphomycosis due to Curvularia lunata (Cochliobolus lunatus) in a cat

A 13-year-old female neutered domestic short-hair cat was presented with chronic progressive vestibular ataxia, lethargy and anorexia. Clinical examination revealed bilateral mucopurulent nasal discharge. Neurological examination revealed obtundation, a right head tilt, ambulatory tetraparesis, generalised vestibular ataxia, decreased postural reactions in all limbs, right Horner's syndrome, spontaneous conjugate jerk rotatory nystagmus and right positional ventral strabismus. Neuroanatomical localisation was observed in the right central vestibular system. Computed tomography revealed a solitary ill-defined contrast-enhancing mass lesion at the level of the right cerebellopontine angle. Cerebrospinal fluid (CSF) analysis revealed mild mononuclear pleocytosis and fungal elements. CSF culture was positive for Curvularia spp. Further tests for underlying diseases were all negative. The cat was treated with antibiotic and antifungal treatment, but it deteriorated rapidly and was euthanased. Necropsy of the brainstem mass lesion revealed pyogranulomatous inflammation. Panfungal polymerase chain reaction (PCR) targeting the internal transcribed spacer (ITS) region and subsequent sequencing identified Curvularia lunata in the formalin fixed brain tissue. This is the first report of brainstem phaeohyphomycosis by Curvularia lunata (Pleosporales) in a cat. In addition, this is the first report among animal and humans where fungal elements of Curvularia lunata were found in the CSF cytology. Opportunistic fungal pathogens should be always considered within the differential diagnoses list in cats with neurological signs and advanced imaging findings compatible with solitary mass lesions in the brain. In feline patients with pyogranulomatous meningoencephalitis and a suspicion of a fungal aetiology, panfungal PCR for the ITS region and sequencing should be performed regardless of the absence of fungal elements in histopathology

Investigation of hypomagnesaemia prevalence and underlying aetiology in a hospitalised cohort of dogs with ionised hypocalcaemia

OBJECTIVES: Calcium is the most abundant mineral in the body and plays a critical role in a wide range of physiological processes. Low concentrations of ionised calcium, the most metabolically available form of calcium, have been linked to an increased risk of adverse clinical outcomes in dogs. Magnesium plays an important role in parathyroid hormone function. The objective of this study was to define the prevalence and aetiology of hypomagnesaemia in a hospitalised cohort of dogs with ionised hypocalcaemia (IHC).METHODS: A total magnesium reference interval was established using serum biochemistry results from 346 clinically healthy dogs. The clinical records of dogs with IHC were reviewed, and concurrent serum magnesium concentrations were recorded alongside clinical signs and underlying aetiology. The prevalence, clinical presentation and aetiology of hypomagnesaemia were examined in the IHC population.RESULTS: Two hundred and ninety-five IHC dogs were identified. Hypomagnesaemia was identified in 22%. Total magnesium concentration was significantly higher in dogs with renal disease. The most common cause of concurrent hypomagnesaemia and IHC was gastrointestinal diseases.CONCLUSION: Low concentrations of serum magnesium occur in approximately one fifth of all dogs with IHC. Further studies are required to clarify the link between magnesium status, IHC and clinical outcome.</p

Multicenter, retrospective determination of the clinical utility of screening tests in dogs with immune-mediated hemolytic anemia in the United Kingdom and Ireland

BACKGROUND: Potential triggers of immune-mediated hemolytic anemia (IMHA) are often identified, but their frequency and the benefit of extensive screening for these to individual dogs is uncertain.OBJECTIVE: To assess the frequency of non-associative IMHA in dogs undergoing screening in Britain and Ireland and identify where specific tests could be beneficial.ANIMALS: Two hundred twenty-two client-owned dogs with IMHA.METHODS: Multicenter, retrospective cohort study of dogs with IMHA. Medical records and blood, urine, imaging, and pathology reports were reviewed. Cases were assessed for associative IMHA, and multivariable analysis was performed to define those.RESULTS: Associative IMHA was present in 73/222 (33%) dogs. Diagnoses included toxic (24/222, 11%); infectious (17/222, 8%); neoplastic (16/222, 7%) and non-infectious inflammatory (13/222, 6%) conditions. A further 102 dogs (46%) had a finding most likely incidental, with no pertinent findings in 47/222 (21%) dogs. Associative IMHA was more likely as patients aged (odds ratio 1.108 per year, 95% CI: 1.012-1.218, p = 0.03).CONCLUSIONS AND CLINICAL IMPORTANCE: The benefit of extensive diagnostic screening and implication of detected abnormalities remains uncertain for individual dogs with IMHA in Britain and Ireland. However, older dogs are more likely to have pertinent findings after a diagnosis of IMHA.</p

Effects of surgery on free and total 25 hydroxyvitamin D concentrations in dogs

BACKGROUND: It is unclear whether a low total 25(OH)D concentration is a cause or consequence of illnesses. To address this knowledge gap, studies measuring free and total 25(OH)D during the evolution and resolution of an inflammatory process are required.OBJECTIVES: Serum total and free 25(OH)D concentrations would transiently decline after cruciate surgery in dogs.ANIMALS: Seventeen client-owned dogs with a spontaneous cranial cruciate ligament rupture (CCLR).METHODS: A longitudinal cohort study involving the measurement of serum concentrations of total and free 25(OH)D, total calcium, creatinine, albumin, phosphate, C-reactive protein and plasma ionized calcium, at 1 day before and a median time of 1 and 60 days after surgical treatment of CCLR.RESULTS: Median serum concentrations of total 25(OH)D before surgery (80.3 nmoL/L [range, 43.5-137.3]) significantly declined immediately after surgery; (64.8 nmoL/L [range, 36.3-116.5] 1 day after surgery, P &lt; .005) before increasing to become nonsignificantly different from concentrations before surgery at day 60 after surgery (median 78.0 nmoL/L [range, 24.2-115.8], P = .14). In contrast, median free 25(OH)D concentrations before surgery (7.6 pg/mL [range, 3.8-12.2]) significantly increased immediately after surgery (9.2 pg/mL [range, 5.2-15.7], P &lt; .05) before declining to become nonsignificantly different from before surgery concentrations at day 60 after surgery (median 6.2 pg/mL [range, 4.0-15.8], P = .37).CONCLUSION AND CLINICAL IMPORTANCE: This study reveals the difficulties of assessing vitamin D status in dogs following elective surgery.</p

Unpublished Mediterranean and Black Sea records of marine alien, cryptogenic, and neonative species

To enrich spatio-temporal information on the distribution of alien, cryptogenic, and neonative species in the Mediterranean and the Black Sea, a collective effort by 173 marine scientists was made to provide unpublished records and make them open access to the scientific community. Through this effort, we collected and harmonized a dataset of 12,649 records. It includes 247 taxa, of which 217 are Animalia, 25 Plantae and 5 Chromista, from 23 countries surrounding the Mediterranean and the Black Sea. Chordata was the most abundant taxonomic group, followed by Arthropoda, Mollusca, and Annelida. In terms of species records, Siganus luridus, Siganus rivulatus, Saurida lessepsianus, Pterois miles, Upeneus moluccensis, Charybdis (Archias) longicollis, and Caulerpa cylindracea were the most numerous. The temporal distribution of the records ranges from 1973 to 2022, with 44% of the records in 2020–2021. Lethrinus borbonicus is reported for the first time in the Mediterranean Sea, while Pomatoschistus quagga, Caulerpa cylindracea, Grateloupia turuturu, and Misophria pallida are first records for the Black Sea; Kapraunia schneideri is recorded for the second time in the Mediterranean and for the first time in Israel; Prionospio depauperata and Pseudonereis anomala are reported for the first time from the Sea of Marmara. Many first country records are also included, namely: Amathia verticillata (Montenegro), Ampithoe valida (Italy), Antithamnion amphigeneum (Greece), Clavelina oblonga (Tunisia and Slovenia), Dendostrea cf. folium (Syria), Epinephelus fasciatus (Tunisia), Ganonema farinosum (Montenegro), Macrorhynchia philippina (Tunisia), Marenzelleria neglecta (Romania), Paratapes textilis (Tunisia), and Botrylloides diegensis (Tunisia)