Verification of the fCAL turbo immunoturbidimetric assay for measurement of the fecal calprotectin concentration in dogs and cats

The concentration of calprotectin in feces (fCal) is a clinically useful marker of chronic gastrointestinal inflammation in humans and dogs. No commercial assay is widely available to measure fCal in small animal medicine, to date. Thus, we verified the immunoturbidimetric fCAL turbo assay (Bühlmann) of fCal for canine and feline fecal extracts by determining linearity, spiking and recovery, and intra-assay and inter-assay variability. We determined RIs, temporal variation over 3 mo, and effect of vaccination and NSAID treatment. Observed:expected (O:E) ratios ($\bar{x}$ ± SD) for serial dilutions of feces were 89–131% (106 ± 9%) in dogs and 77–122% (100 ± 12%) in cats. For spiking and recovery, the O:E ratios were 90–118% (102 ± 11%) in dogs and 83–235% (129 ± 42%) in cats. Intra- and inter-assay CVs for canine samples were ≤19% and ≤7%, and for feline samples ≤22% and ≤21%. Single-sample RIs were <41 μg/g for dogs and <64 μg/g for cats. With low reciprocal individuality indices, using population-based fCal RIs is appropriate, and moderate fCal changes between measurements (dogs 44.0%; cats: 43.2%) are considered relevant. Cats had significant (but unlikely relevant) fCal increases post-vaccination. Despite individual fCal spikes, no differences were seen during NSAID treatment. The fCAL turbidimetric assay is linear, precise, reproducible, and sufficiently accurate for measuring fCal in dogs and cats. Careful interpretation of fCal concentrations is warranted in both species during the peri-vaccination period and for some patients receiving NSAID treatment.     

Changes in the blood lactate dehydrogenase measurements in canines and felines following the international standardization of the assay method

Lactate dehydrogenase (LDH) in blood is measured using the Japanese Society of Clinical Chemistry (JSCC) method in Japan and the International Federation of Clinical Chemistry (IFCC) method in other countries. In human clinical practice, the IFCC method replaced the JSCC method due to international standardization. Moreover, veterinary LDH measurement will also eventually shift to the IFCC method. However, the relationship between the IFCC and JSCC methods for LDH in various animals and whether they can be equated or not have not yet been investigated. This study aimed to present the changes in measurements in canines and felines after switching to the IFCC method. The plasma LDH activity of canines (N=177) and felines (N=115), who visited a secondary care veterinary clinic, was measured using the JSCC and IFCC methods. The IFCC/JSCC ratio was <1.0 in 85% of canines and 88% of felines, indicating that the IFCC method tended to show lower values than the JSCC method, presumably because LDH5 is dominant among the LDH isozymes in canines and felines. The increase in the systematic errors of both assays was in the high value range, with some samples exceeding the error tolerance from near the upper end of the reference range. When switching to the IFCC method for LDH measurement in canines and felines, each institution should consider whether the reference range and clinical diagnostic values established by the JSCC method are appropriate for continued use.     

Comparison of three ultrasound guided approaches to the lumbar plexus in dogs: a cadaveric study

ObjectiveTo assess the accuracy of contrast material injection and the dispersion of injectate following ultrasound guided injections at the level of L6 and L7, in canine cadavers.Study designProspective, randomized, experimental study.AnimalsTwenty nine mixed breed canine cadavers (28.9 ± 6.0 kg).MethodsThree ultrasound-guided approaches to the lumbar plexus (LP) were compared: 1) a dorsal pre-iliac approach at the level of L6; 2) a lateral paravertebral approach at mid-L6; and 3) a lateral paravertebral approach at mid-L7. An isovolumic mixture of iodine-based contrast with new methylene blue (0.1 mL kg⁻¹) was injected bilaterally in the juxta-foraminal region along the L6 or L7 nerve root. Computed tomography was performed followed by segmentation and 3D reconstruction of the lumbar spine and contrast material volumes using dedicated software. Distances between contrast material and the fifth through seventh lumbar foraminae, and length of femoral (FN) and obturator (ON) nerve staining were measured and compared between approaches (p < 0.05).ResultsInjectate moved cranial and caudal to the site of injection, and dispersed into an ovoid shape between the quadratus lumborum, iliopsoas and psoas minor muscles. Injections at L7 resulted in significantly closer contrast proximity to the L6 and L7 foraminae (p < 0.001). Femoral nerve staining was similar for all approaches, ON staining was more consistent after L7 injections (p < 0.001).Conclusion and clinical relevanceAn ultrasound-guided lateral paravertebral approach to the LP proved very practical and accurate, with easy visualization of the plexus and associated nerves. To ensure that the ON is covered by injectate, an approach at the level of L7 is recommended. Further studies are necessary to determine if this correlates with clinically effective local anesthesia.     

Increased canine pancreatic lipase immunoreactivity (cPLI) and 1,2-o-dilauryl-rac-glycero-3-glutaric acid-(6′-methylresorufin) ester (DGGR) lipase in dogs with evidence of portal hypertension and normal pancreatic histology: a pilot study

The clinical presentations of both liver disease and pancreatitis are nonspecific and overlapping, which may cause difficulty in diagnosis. In our retrospective pilot study, we assessed whether dogs with evidence of portal hypertension and absence of pancreatitis on pancreatic histology have increases in canine pancreatic lipase immunoreactivity (cPLI) and 1,2-o-dilauryl-rac-glycero-3-glutaric acid-(6′-methylresorufin) ester (DGGR) lipase. We included dogs that had been presented between 2008 and 2019 if they had normal pancreatic histology, histologically confirmed hepatopathy, and if canine pancreas-specific lipase (Spec cPL; Idexx) or DGGR lipase had been measured. Only dogs with portal hypertension were included. Six dogs fulfilled the inclusion criteria. Four of 6 and 2 of 6 dogs had Spec cPL and DGGR lipase exceeding the upper reference limit, respectively. From the 4 dogs with increased Spec cPL, 2 had concentrations of 200–400 µg/L and 2 had concentrations ≥ 400 µg/L. Our results suggest that canine portal hypertension might lead to increased Spec cPL and DGGR lipase values in the absence of pancreatitis on histology. Until more evidence in a larger number of dogs with portal hypertension is available, both tests should be interpreted cautiously in the presence of portal hypertension.     

Infective endocarditis of the aortic valve in a Border collie dog with patent ductus arteriosus

Infective endocarditis (IE) in dogs with cardiac shunts has not been reported previously. However, we encountered a dog with concurrent patent ductus arteriosus (PDA) and IE. The dog was a 1-year-old, 13.9-kg female Border collie and presented with anorexia, weight loss, pyrexia (40.4°C) and lameness. A continuous murmur with maximal intensity over the left heart base (Levine 5/6) was detected on auscultation. Echocardiography revealed a PDA and severe aortic stenosis (AS) caused by aortic-valve vegetative lesions. Corynebacterium spp. and Bacillus subtilis were isolated from blood cultures. The dog responded to aggressive antibiotic therapy, and the PDA was subsequently surgically corrected. After a series of treatments, the dog showed long-term improvement in clinical status.     

Ventrolateral temporal lobectomy in normal dogs as a counterpart to human anterior temporal lobectomy: a preliminary study on the surgical procedure and complications

Anterior temporal lobectomy (ATL) is a surgical procedure for drug-resistant mesial temporal lobe epilepsy that is commonly performed in human medicine. The purpose of this study was to determine whether ATL-like surgery, i.e., removal of the amygdala and hippocampal head, is possible in dogs, and to investigate its safety and postoperative complications. Eight healthy beagles underwent ATL-like surgery and were observed for 3 months postoperatively. Samples from the surgically resected tissues and postmortem brain were evaluated pathologically. The surgical survival rate was 62.5%. The major postoperative complications were visual impairment, temporal muscle atrophy on the operative side, and a postoperative acute symptomatic seizure. Due to the anatomical differences between dogs and humans, the surgically resected area to approach the medial temporal structures in dogs was the ventrolateral part of the temporal lobe. Therefore, the ATL-like surgery described in this study was named “ventrolateral temporal lobectomy” (VTL). This study is the first report of temporal lobectomy including amygdalohippocampectomy in veterinary medicine and demonstrates its feasibility. Although it requires some degree of skill, VTL could be a treatment option for canine drug-resistant epilepsy and lesions in the mesial temporal lobe.     

An Earlier Uterine Environment Favors the In Vivo Development of Fresh Pig Morulae and Blastocysts Transferred by a Nonsurgical Deep-uterine Method

This study aimed to evaluate the effect of recipient-donor estrous cycle synchrony on recipient reproductive performance after nonsurgical deep-uterine (NsDU) embryo transfer (ET). The transfers (N=132) were conducted in recipients sows that started estrus 24 h before (–24 h; N=9) or 0 h (synchronous; N=31), 24 h (+24 h; N=74) or 48 h (+48 h; N=18) after the donors. A total of 30 day 5 morulae or day 6 blastocysts (day 0=onset of estrus) were transferred per recipient. The highest farrowing rates (FRs) were achieved when estrus appeared in recipients 24 h later than that in the donors (81.1%), regardless of the embryonic stage used for the transfers. The FR notably decreased (P<0.05) when recipients were –24 h asynchronous (0%), synchronous (61.3%) or +48 h asynchronous (50%) relative to the donors. No differences in litter size (LS) and piglet birth weights were observed among the synchronous and +24 h or +48 h asynchronous groups. While a +24 h asynchronous recipient was suitable for transfers performed with either morulae (FR, 74.3%; LS, 9.2 ± 0.6 piglets) or blastocysts (FR, 84.6%; LS, 9.8 ± 0.6 piglets), a + 48 h asynchronous recipient was adequate for blastocysts (FR, 87.5%; LS, 10.4 ± 0.7 piglets) but not for morulae (FR, 30.0%; LS, 7.3 ± 2.3 piglets). In conclusion, this study confirms the effectiveness of the NsDU-ET technology and shows that porcine embryos tolerate better a less advanced uterine environment if they are nonsurgically transferred deep into the uterine horn.     

Identification and preservation of the parathyroid gland during total thyroidectomy in dogs with bilateral thyroid carcinoma: a report of six cases

Simultaneous removal of bilateral thyroid tumors was performed while preserving the parathyroid gland in six dogs. At least one external parathyroid gland was identified in all dogs. In five cases, the external parathyroid gland and its blood supply were preserved intact. In one dog, the vessels supplying the external parathyroid gland had been invaded by the tumor, and the gland was thus removed and reimplanted into the sternohyoid muscle. That dog required postoperative treatment with oral calcium gluconate and vitamin D₃. Local tumor recurrence was not observed in any of the cases. The mean survival time was 920 days. We found that the external parathyroid gland could be identified and preserved in most dogs undergoing total thyroidectomy.     

A Pilot Study to Assess the Feasibility of a Submaximal Exercise Test to Measure Individual Response to Cardiac Medication in Dogs with Acquired Heart Failure

Exercise testing is not commonly used in canine medicine because of several limitations. The aim of this study was to investigate the suitability of a treadmill test to measure the exercise capacity of untrained canine cardiac patients and to measure some biological parameters that might reflect the tolerance of dogs with heart failure to submaximal exercise. The exercise capacity of seven dogs with naturally occurring heart failure was evaluated before the institution of cardiac medication and 7 days after the beginning of the study. An additional re-examination was requested after 28 days. The exercise test was performed on a motorized treadmill at three different speeds (0.5 m/s, 1.0 m/s and 1.5 m/s). The following parameters were measured at the end of each stage and after 20 min recovery: heart rate, rectal temperature, glucose, lactate, aspartate aminotransferase, creatine kinase, Pvo ₂, Pvco ₂, pH, haematocrit, bicarbonate, sodium, potassium and chloride. Serum cardiac troponin-I was also measured at the beginning of the test and at the end of the recovery period. Owners’ perception reflected the ability of their dogs to exercise on the treadmill. Lactate level increased noticeably with the intensity of the exercise test, and its variation coincided with different exercise tolerance observed by the owners. Heart rate seemed to follow a similar trend in the few dogs presented in sinus rhythm. None of the remaining parameters appeared to be sensitive indicators of activity level in the dogs used in this study. The treadmill exercise test in dogs with acquired heart failure is feasible and might provide useful information for assessing individual response to cardiac medication. Lactate and heart rate seemed to reflect individual levels of exercise tolerance, although further studies are necessary to confirm the reliability and repeatability of this test.     

Treatment with a High Dose Combination of PMSG/hCG Preparation of Mares Clinically Diagnosed with Ovarian Quiescence during the Breeding Season (Investigation from 1975 to 2000)

A total of 88 thoroughbred mares were diagnosed with clinical ovarian quiescence and subjected to four treatment regimens. Using PMSG, hCG or combinations of both. A high dose combination of 5,000IU PMSG with 5,000IU hCG showed significantly higher rates of marked estrus and ovulation induction (P<0.01) as well as conception rates (P<0.05). In the present study, the administration of a high-dose combination of PMSG with hCG was shown to be an effective treatment of ovarian quiescence in light mares.     

Establishment of a Conditional Transgenic System Using the 2A Peptide in the Female Mouse Germline

Transgenic mice are essential research tools in developmental biology studies. The 2A peptide allows multiple genes to be expressed simultaneously at comparable levels in somatic cells, but there are no reports of it being used successfully in germ cells. We constructed a Cre/loxP-based conditional vector containing the 2A peptide to significantly enhance the expression of a reporter and target gene from a constitutive promoter in oocytes. Mice with a transgene insertion containing the chicken β-actin promoter, floxed EGFP-polyA cassette, mCherry reporter, 2A peptide and target gene DNA methyltransferase 3A2 (Dnmt3a2) were crossed with TNAP- or Vasa-Cre mice to produce offspring, in which mCherry and DNMT3A2 proteins were highly expressed in oocytes upon Cre-mediated removal of EGFP-polyA. This novel transgenic mouse line based on the 2A expression system can serve as a useful tool for examining gene function during oogenesis.     

Business intelligence tools to optimize the appropriateness of the diagnostic process for clinical and epidemiologic purposes in a multicenter veterinary pathology service

Laboratory tests provide essential support to the veterinary practitioner, and their use has grown exponentially. This growth is the result of several factors, such as the eradication of historical diseases, the occurrence of multifactorial diseases, and the obligation to control endemic and epidemic diseases. However, the introduction of novel techniques is counterbalanced by economic constraints, and the establishment of evidence- and consensus-based guidelines is essential to support the pathologist. Therefore, we developed standardized protocols, categorized by species, type of production, age, and syndrome at the Istituto Zooprofilattico Sperimentale delle Venezie (IZSVe), a multicenter institution for animal health and food safety. We have 72 protocols in use for livestock, poultry, and pets, categorized as, for example, “bovine enteric calf”, “rabbit respiratory”, “broiler articular”. Each protocol consists of a panel of tests, divided into ‘mandatory’ and ‘ancillary’, to be selected by the pathologist in order to reach the final diagnosis. After autopsy, the case is categorized into a specific syndrome, subsequently referred to as a syndrome-specific panel of analyses. The activity of the laboratories is monitored through a web-based dynamic reporting system developed using a business intelligence product (QlikView) connected to the laboratory information management system (IZILAB). On a daily basis, reports become available at general, laboratory, and case levels, and are updated as needed. The reporting system highlights epidemiologic variations in the field and allows verification of compliance with the protocols within the organization. The diagnostic protocols are revised annually to increase system efficiency and to address stakeholder requests.     

Alteration of the Endometrial EGF Profile as a Potential Mechanism Connecting the Alterations in the Ovarian Steroid Hormone Profile to Embryonic Loss in Repeat Breeders and High-producing Cows

Poor reproductive efficiency is a worldwide problem that has affected the dairy industry during the last several decades. In an attempt to explain the changes in reproductive physiology caused by high milk production, a model of elevated steroid metabolism in lactating dairy cows has been proposed. A slow increase in levels and low peak levels of estradiol (E₂) and progesterone (P₄) characterize endocrine changes in high producing cows. Similar changes have been reported in the repeat breeder cows. The abnormal changes in E₂ and P₄ concentrations of these cows may cause an improper uterine environment due to disturbed expression of growth factors and cytokines in the endometrium. This review focuses on the alteration in epidermal growth factor (EGF) profile in the endometrium during the estrous cycle. The normal cow has two peaks of EGF concentrations on days 2–4 and 13–14. Low concentrations of EGF on these days distinguished both high-producing and repeat breeder cows from normal cows. Alteration of the EGF profile could be found in 70 and 40% of the repeat breeder and high-producing cows, respectively. Treatment with a high dose of estradiol benzoate and an intravaginal progesterone-releasing device restored the normal EGF profile in about 70% of the affected cows. The cows having a normal EGF profile after treatment showed a higher pregnancy rate than the cows with the altered profile. Further studies to understand the etiology of the alteration in the EGF profile are needed to develop another treatment option and preventive management for this problem.     

Long-term postsurgical outcomes of mast cell tumors resected with a margin proportional to the tumor diameter in 23 dogs

We evaluated the postsurgical outcomes of cutaneous or subcutaneous mast cell tumors (MCTs, n=25) in 23 dogs, resected with lateral surgical margins proportional to the widest tumor diameter, including at least one facial plane. The tumor diameter range was 0.3–2.6 cm (median: 0.9 cm), and all were histologically diagnosed as Kiupel’s low-grade MCT. Resection was histologically complete in 20, close (deep margin) in three, and incomplete (deep margin) in two. No dogs developed local recurrence at the site of initial surgery during follow-up of 161–2,219 days (median: 976 days). These results suggest that resection of low-grade, relatively small MCTs with surgical margins proportional to the tumor diameter is a practical procedure with high success rates.     

A subjective descriptive study of the warm‐up and turn to a fence,approach, take‐off,suspension, landing and move‐off in 10 showjumpers

There is limited knowledge about causes of musculoskeletal injury in showjumpers. The objectives were to describe features of the turn, approach and jump in a group of experienced showjumpers believed by their riders to be sound, to relate these findings to clinical findings, and to identify features that may predispose to injury. Ten experienced showjumpers in normal competition training jumped an upright and a parallel fence four times off the left and right reins respectively, after a rider‐defined period of warm‐up. Real‐time and high‐speed motion capture was undertaken. Detailed subjective assessment of the gait was performed during the warm‐up, on the turn and approach to the jump and all phases of the jump. Six horses had thoracolumbar pain or epaxial muscle tension. Six horses exhibited a poor‐quality canter. The mean duration of warm‐up was 7 min (range 5–10 min). All horses had lean of the trunk and hindlimbs >45° on the turn. The inside hindlimb was placed in front of the outside hindlimb in 75% of the turns. Sideways oscillations of the hocks during stance were seen in all horses on the turn and on the straight approach in five horses. The inside hindlimb had greater magnitude of oscillation than the outside hindlimb on the turns. Repeated asynchronous push‐off with the hindlimbs at take‐off was seen in three horses. The hindlimbs drifted to the left or right during the ascent‐phase of suspension in four horses, reflecting asymmetrical push‐off. Only two horses landed consistently with the correct forelimb leading relative to the direction in which the horse had to turn after the fence. Four horses landed seven (n = 2) or eight (n = 2) times each with the left (n = 2) and right (n = 2) forelimbs respectively. Repetitive overload through asymmetrical use of the left and right canter leads, inadequate warm‐up, and limb instability could potentially predispose to injury.