Vet. Sci., Volume 8, Issue 3 (March 2021) – 18 articles

Feline squamous cell carcinoma (SCC) is currently treated with surgery, radiation therapy and electrochemotherapy (ECT). Both the efficacy and/or safety of ECT were evaluated as a sole therapy with bleomycin to treat feline nasal planum SCC (npSCC). Sixty-one cats were enrolled. Local treatment response was evaluated as complete remission (CR), partial remission (PR) or stable disease (SD). Recurrence rate (RR), disease-free interval (DFI) and progression free survival (PFS) were calculated. A six-point scale was used for ECT toxicity. The median tumor size was 1.5 cm. CR was achieved in 65.6% of cases, PR in 31.1% and SD in 3.3%. The overall response rate was 96.7%, RR was 22.5%, median DFI was 136 days, and median PFS was 65.5 days. ECT toxicity was ≤2 in 51% of cats. Tumor recurrence/progression (p = 0.014) and local treatment response (PR: p < 0.001; SD: p < 0.001) influenced survival time. Cats with toxicity >2 showed a higher probability of tumor recurrence/progression. Tumor-related death was higher in cats with PR (p < 0.001) and recurrence/progression (p = 0.002), in ECT treatment with 1 Hz (p = 0.035) and 1200 V/cm (p = 0.011) or 1300 V/cm (p = 0.016). Tumor size influenced local treatment response (p = 0.008) and toxicity (p < 0.001). ECT is an effective treatment for feline npSCCs and should be considered as the first-line procedure for low-stage tumors. Full article

Saccharomyces cerevisiae is a yeast strain often used to improve the feed quality of ruminants. However, S. cerevisiae has limited capacity to provide biomass when inoculated with carbon sources and a low ability to produce cellulase enzymes. Here, we hypothesized that yeast in the rumen produces a large amount of biomass and could release cellulase enzymes to break down fiber content. Therefore, the aim of this study was to screen, isolate and identify yeast from the rumen fluids of Holstein Friesian steers and measure the efficiency of biomass production and cellulase activity. A fermentation medium containing sugarcane molasses as a carbon source and urea as a nitrogen source was optimized. Two fistulated–crossbred Holstein Friesian steers averaging 350 ± 20 kg body weight were used to screen and isolate the ruminal yeast. Two experiments were designed: First, a 12 × 3 × 3 factorial was used in a completely randomized design to determine biomass and carboxymethyl cellulase activity. Factor A was the isolated yeast and S. cerevisiae. Factor B was sugarcane molasses (M) concentration. Factor C was urea (U) concentration. In the second experiment, potential yeasts were selected, identified, and analyzed for 7 × 4 factorial use in a completely randomized design. Factor A was the incubation times. Factor B was the isolated yeast strains, including codes H-Khon Kaen University (KKU) 20 (as P. kudriavzevii-KKU20), I-KKU20 (C. tropicalis-KKU20), and C-KKU20 (as Galactomyces sp.-KKU20). Isolation was imposed under aerobic conditions, resulting in a total of 11 different colonies. Two appearances of colonies including asymmetric colonies of isolated yeast (indicated as A, B, C, E, and J) and ovoid colonies (coded as D, F, G, H, I, and K) were noted. Isolated yeast from the rumen capable of providing a high amount of biomass when inoculant consisted of the molasses 15% + urea 3% (M15 + U3), molasses 25% + urea 1% (M25 + U1), molasses 25% + urea 3% (M25 + U3), and molasses 25% + urea 5% (M25 + U5) when compared to the other media solution (p < 0.01). In addition, 11 isolated biomass-producing yeasts were found in the media solution of M25 + U1. There were 4 isolates cellulase producing yeasts discovered in the media solution of M25 + U1 and M25 + U5 whereas molasses 5% + urea 1% (M5 + U1), molasses 5% + urea 3% (M5 + U3), molasses 5% + urea 5% (M5 + U5), molasses 15% + urea 1% (M15 + U1), molasses 15% + urea 3% (M5 + U3), and M25 + U3 were found with 2, 3, 1, 2, 1, and 2 isolates, respectively. Ruminal yeast strains H-KKU20, I-KKU20, and C-KKU20 were selected for their ability to produce biomass. Identification of isolates H-KKU20 and I-KKU20 revealed that those isolates belonged to Pichia kudriavzevii-KKU20 and Candida tropicalis-KKU20 while C-KKU20 was identified as Galactomyces sp.-KKU20. Two strains provided maximum cell growth: P. kudriavzevii-KKU20 (9.78 and 10.02 Log cell/mL) and C. tropicalis-KKU20 (9.53 and 9.6 Log cells/mL) at 60 and 72 h of incubation time, respectively. The highest ethanol production was observed in S. cerevisiae at 76.4, 77.8, 78.5, and 78.6 g/L at 36, 48, 60, and 72 h of incubation time, respectively (p < 0.01). The P. kudriavzevii-KKU20 yielded the least reducing sugar at about 30.6 and 29.8 g/L at 60 and 72 h of incubation time, respectively. The screening and isolation of yeasts from rumen fluids resulted in 11 different yeasts being obtained. The potential yeasts discovered in the rumen fluid of cattle were Pichia kudriavzevii-KKU20, Candida tropicalis-KKU20, and Galactomyces sp.-KKU20. P. kudriavzevii-KKU20 had higher results than the other yeasts in terms of biomass production, cellulase enzyme activity, and cell number. Full article

Acute pancreatitis is an acute inflammatory process in the pancreas that is common in dogs. This study was designed to compare cytokines between healthy dogs and dogs with suspected acute pancreatitis. For the canine cytokine antibody array, three healthy dogs and three dogs with suspected acute pancreatitis were included. Interleukin (IL)-2, IL-6, IL-10, GM-CSF, and TNF-α were not detected in either group based on the results. Conversely, IL-8 (p = 0.035), Monocyte Chemoattractant Protein-1 (MCP)-1 (p = 0.0138), Receptor for Advanced Glycation Endproducts (RAGE) (p = 0.0079), and stem cell factor (SCF) (p = 0.034) were significantly increased in dogs with suspected acute pancreatitis. However, vascular endothelial growth factor (VEGF) (p = 0.6971) did not differ significantly between groups. For the canine serum Enzyme-Linked Immunosorbent Assay (ELISA), eight healthy dogs and eight dogs with suspected acute pancreatitis were included. ELISA revealed that IL-8 (p < 0.0001), MCP-1 (p < 0.0001), RAGE (p = 0.006), and SCF (p = 0.0002) were all significantly upregulated in the experimental group. We confirmed multiple patterns of cytokines in suspected acute pancreatitis of dogs via canine cytokine antibody array using a small quantity of serum. After this procedure, we reevaluated the cytokines, which were significantly increased in dogs with suspected acute pancreatitis, by ELISA, with more samples. Through this study, we confirmed that MCP-1, RAGE, and SCF were newly suggested factors in dogs with suspected acute pancreatitis. Full article

Male infertility is an emerging problem in both humans and animals, and the knowledge of its causes is the first step to identifying new diagnostic and therapeutic strategies. In humans, alteration of sperm DNA methylation have been related to poor quality semen, impaired seminal parameters, azoospermia and reduced fertility. Although semen analysis is routinely used to evaluate the male reproductive potential in the canine species, no authors have attempted to relate semen characteristics to the sperm global DNA methylation (SGDM). The aim of this study was to evaluate the SGDM level in healthy dogs and to correlate it with semen parameters that are currently used in dog semen analyses. Conventional and unconventional (sperm DNA fragmentation and SGDM) seminal parameters of thirty dogs from different breeds were evaluated. A positive correlation was found between SGDM and sperm concentration (r = 0.41; p < 0.05), and total sperm count (r = 0.61; p < 0.001); SGDM was significantly lower in oligozoospermic vs non-oligozoospermic dogs (4.3% vs. 8.7%; p < 0.005). Our findings suggest that SGDM levels are related to conventional seminal parameters, and could be used as a marker of testis function and spermatogenesis in dogs. Full article

DBA/2 mice are a well-known animal model for hearing loss developed due to intrinsic properties of these animals. However, results on the phenotype of hearing loss in DBA/2 mice have been mainly reported at an early stage in mice aged ≤7 weeks. Instead, the present study evaluated the hearing ability at 5, 13, and 34 weeks of age using DBA/2korl mice. Auditory brainstem response test was performed at 8–32 KHz at 5, 13, and 34 weeks of age, and hearing loss was confirmed to be induced in a time-dependent manner. In addition, histopathological evaluation at the same age confirmed the morphological damage of the cochlea. The findings presented herein are the results of the long-term observation of the phenotype of hearing loss in DBA/2 mice and can be useful in studies related to aging-dependent hearing loss. Full article

The anaerobic bacterium Cutibacterium acnes has been increasingly linked to the development of degenerative disc disease (DDD), although causality is yet to be conclusively proven. To better study how this organism could contribute to the aetiology of DDD, improved animal models that are more reflective of human disc anatomy, biology and mechanical properties are required. Against this background, our proof-of concept study aimed to be the first demonstration that C. acnes could be safely administered percutaneously into sheep intervertebral discs (IVDs) for in vivo study. Following our protocol, two sheep were successfully injected with a strain of C. acnes (8.3 × 10⁶ CFU/disc) previously recovered from a human degenerative disc. No adverse reactions were noted, and at one-month post inoculation all triplicate infected discs in our first animal grew C. acnes, albeit at a reduced load (5.12 × 10⁴ to 6.67 × 10⁴ CFU/disc). At six months, no growth was detected in discs from our second animal indicating bacterial clearance. This pilot study has demonstrated the feasibility of safe percutaneous injection of C. acnes into sheep IVDs under fluoroscopic guidance. The design of follow-up sheep studies to investigate the potential of C. acnes to drive pathological changes within infected discs should now be pursued. Full article

Infectious bronchitis virus (IBV) is a major economic problem in commercial chicken farms with acute multiple-system infection, especially in respiratory and urogenital systems. A live-attenuated and killed vaccine is currently immunized to control IBV infection; however, repeated outbreaks occur in both unvaccinated and vaccinated birds due to the choice of inadequate vaccine candidates and continuous emergence of novel infectious bronchitis (IB) variants and failure of vaccination. However, similar clinical signs were shown in different respiratory diseases that are essential to improving the diagnostic assay to detect IBV infections. Various risk factors involved in the failure of IB vaccination, such as various routes of application of vaccination, the interval between vaccinations, and challenge with various possible immunosuppression of birds are reviewed. The review article also highlights and updates factors affecting the diagnosis of IBV disease in the poultry industry with differential diagnosis to find the nature of infections compared with non-IBV diseases. Therefore, it is essential to monitor the common reasons for failed IBV vaccinations with preventive action, and proper diagnostic facilities for identifying the infective stage, leading to earlier control and reduced economic losses from IBV disease. Full article

The highly toxic Aflatoxin M1 (AFM1) is most often detected in milk using an Enzyme-Linked-Immunosorbent Assay (ELISA) for screening purposes, while High-Performance Liquid Chromatography with Fluorescence Detector (HPLC-FL) is the reference method used for confirmation. The aim of the present study was the comparison between three commercially available ELISA kits and a newly developed HPLC-FL method for the determination of the AFM1 in milk samples. The developed HPLC-FL method was validated for the AFM1 and Aflatoxin M2 (AFM2), determining the accuracy, precision, linearity, decision limit, and detection capability with fairly good results. All three ELISA kits were also validated and showed equally good performance with high recovery rates. Moreover, the Limit Of Detection (LOD) and Limit Of Quantification (LOQ) values were found to be significantly lower than the Maximum Residue Limit (MRL) (50 ng kg⁻¹). After the evaluation of all three commercial kits, the ELISA kit with the optimum performance along with the HPLC method was used for the determination of AFM1 in raw cow’s, goat’s, and sheep’s milk samples (396) obtained from producers in different regions of Greece. The evaluation of both methods showed that this ELISA kit could be considered as a faster and equally reliable alternative method to HPLC in routine analysis for the determination of AFM1 in milk. Full article

The aim or this study was to determine the relationship between non-esterified fatty acids and biomarkers from an automatic milking system (AMS). Fresh dairy cows (n = 102) were selected and milked in Lely Astronaut^® A3 milking robots. The rumination time (RT), body weight (BW), milk content and composition parameters, milk fat/protein ratio (F/P), and milk electrical conductivity were registered by the same milking robots. For examining non-esterified fatty acids (NEFAs), blood samples were acquired from cows in the dry period. According to the NEFA concentrations, all cows were divided into two groups: Group I, with <0.300 mEq/L (n = 66), and Group II, with ≥0.300 mEq/L (n = 36). Albumin (ALB), aspartate aminotransferase (AST), gamma-glutamyl transferase (GGT), and cortisol concentrations were also analyzed once a week up to 30 days in milking. The study revealed that the cows in Group I had higher concentrations of ALB, cortisol, and GGT, but the average concentration of AST was lower. In Group 1, the milk F/P was higher, but the milk yield was lower. We hypothesize that biomarkers from AMS could help in the early diagnosis of metabolic diseases after calving or to control negative energy balance before calving. Full article

The morphological characteristics of the largest lymphatic vessels and lymph nodes of the body have been described through ultrasonography, although food and gas in the gastrointestinal tract can often have negative effects on the response of small abdominal structures. The aim of the study was to describe the size of normal abdominal lymph nodes (ALs) in dogs affected by disease, not including lymphadenomegaly or lymphadenopathy, and divided according to body weight and age. The ALs studied included the jejunal, medial iliac, portal, gastric, splenic, and pancreaticoduodenal lymph nodes. Statistical correlation considering body weight and age as continuous variables showed that all measurements of the ALs increased according to body weight changes (p < 0.01). The most reliable values were the volume measurements (p < 0.001) compared to the length, thickness, and width. Mixed results emerged from a comparison of weight categories and age; only the jejunal lymph nodes showed a significant correlation (p < 0.05). Other characteristics (shape, attenuation, and enhancement) are subsequently reported. The resulting data can be used to categorize CT measurements of normal ALs displayed based on the body weight and age of the subjects. This study aimed to propose a new parameter of normalcy that may serve as a reference for the evaluation of infectious or neoplastic events. Full article

This report describes the first clinical case, to our knowledge, of a dog with polyglandular deficiency syndrome with diabetes mellitus and hypoadrenocorticism. A six-year-old female Cavalier King Charles Spaniel presented with a history of lethargy and appetite loss. The dog was diagnosed with diabetic ketoacidosis based on hyperglycemia and renal glucose and ketone body loss. The dog’s condition improved on intensive treatment of diabetes mellitus; daily subcutaneous insulin detemir injection maintained an appropriate blood glucose level over half a year. However, the dog’s body weight gradually decreased from day 207, and on day 501, it presented with a decreased appetite; the precise cause could not be determined. Based on mild hyponatremia and hyperkalemia, hypoadrenocorticism was suggested; the diagnosis was made using an adrenocorticotropic hormone stimulation test. Daily fludrocortisone with low-dose prednisolone oral administration resulted in poor recovery of the blood chemistry abnormalities; however, monthly desoxycorticosterone pivalate (DOCP) subcutaneous injection with daily low-dose prednisolone oral administration helped in the significant recovery of the abnormalities. Therefore, clinicians should consider the possibility of coexistence of hypoadrenocorticism in dogs with diabetes mellitus presenting with undifferentiated weight loss. Additionally, DOCP (not fludrocortisone) may be useful in treating dogs with diabetes mellitus complicated with hypoadrenocorticism. Full article

Nematodes are an important cause of disease and loss of performance in horses. Changes in the parasitic fauna of horses have occurred in the past few decades, making cyathostomins the major parasites in adult horses, while large strongyles have become less prevalent. Parascaris spp. remains the most important parasite infecting foals and weanlings. Anthelmintic resistance is highly prevalent in cyathostomins and Parascaris spp. worldwide and it must be factored into treatment decisions. To assess anthelmintic efficacy in Northern Italy, we sampled 215 horses from 17 sport and horse-breeding farms. Fecal egg count reduction tests (FECRT) were used to assess anthelmintic efficacy. Copromicroscopic analysis was performed using MiniFLOTAC before treatment with fenbendazole, pyrantel pamoate or ivermectin, and repeated 14 days post-treatment. Strongyle-type eggs were detected in 66.91% of horses (CI95% 61.40–73.79%), while Parascaris spp. was detected in 2.79% (CI95% 1.94–5.95%). Reduced efficacy against cyathostomins was observed for fenbendazole in 55.56% of the treated animals (CI95% 41.18–69.06%), and for pyrantel pamoate in 75% of animals (CI95% 30.06–95.44%). Ground-based actions must be set in place to promote the uptake of state-of-the-art worm control plans that will prevent clinical disease while minimizing the selection pressure of resistant parasites. Full article

Laboratory tests can be altered in cardiovascular diseases and the investigation of specific tests or biomarkers may provide additional information about myocardial damage. Traditional laboratory tests, such as cell blood count, serum biochemistry, and coagulation, can be useful in investigating patients, but are not specific. However, markers like Troponin and Natriuretic Peptides may possibly furnish further data on myocardium damage and can be used in both studying and monitoring cats with cardiac disease. Moreover, the evaluation of the thyroid profile is very important as hyperthyroid cats concomitant cardiovascular diseases are very common and they can also be a direct consequence of endocrinopathy. The purpose of this manuscript is to provide the widest possible overview of what is present in the literature about the feline clinical pathology of heart diseases through a rational division of the main alterations of traditional tests and biomarkers. Full article

Corrective osteotomy has been applied to realign and stabilize the bones of dogs with lameness. However, corrective osteotomy for angular deformities requires substantial surgical experience for planning and performing accurate osteotomy. Three-dimensional printed patient-specific guides (3D-PSGs) were developed to overcome perioperative difficulties. In addition, novices can easily use these guides for performing accurate corrective osteotomy. We compared the postoperative results of corrective osteotomy accuracy when using 3D-PSGs in dogs between novice and experienced surgeons. We included eight dogs who underwent corrective osteotomy: three angular deformities of the radius and ulna, three distal femoral osteotomies, one center of rotational angle-based leveling osteotomy, and one corrective osteotomy with stifle arthrodesis. All processes, including 3D bone modeling, production of PSGs, and rehearsal surgery were carried out with computer-aided design software and a 3D-printed bone model. Pre- and postoperative positions following 3D reconstruction were evaluated by radiographs using the 2D/3D registration technique. All patients showed clinical improvement with satisfactory alignment and position. Postoperative accuracy evaluation revealed no significant difference between novice and experienced surgeons. PSGs are thought to be useful for novice surgeons to accurately perform corrective osteotomy in dogs without complications. Full article

We conducted a survey of canine microfilaraemia in 768 dogs in Chanthaburi, Samut Sakhon, and Narathiwat provinces of Thailand using a novel semi-automated, microfluidic device that is easy and rapid to perform. Microfilariae species were identified using High Resolution Melting real-time PCR (HRM real-time PCR). The prevalence of canine microfilaremia was 16.2% (45/278) in Chanthaburi and 5.5% (12/217) in Samut Sakhon. The prevalence of canine microfilaremia in Narathiwat was 22.7% (67/273). Brugia pahangi and Dirofilaria immitis were the predominant species of filariae found in the infected dogs from Chanthaburi and Narathiwat, respectively. The low prevalence of canine microfilaremia of Samut Sakhon may reflect the success of the Soi Dog foundation’s efforts and the establishment of veterinary control programs. An effective disease control and prevention strategies is needed in Chanthaburi and Narathiwat to reduce the risks of zoonotic transmission of the parasites. An appropriate drug treatment should be given to infected dogs and prophylactic drugs are suggested to be given to dogs age ≤1-year-old to prevent filarial infection. The novel microfluidic device could be implemented for surveillance of filariae infection in other animals. Full article

Visceral hemangiosarcoma (HSA) is one of the more frequent cancers in dogs and has a high metastatic rate and poor prognosis, as clinical signs only become apparent in advanced stages of tumor development. In order to improve early and differential diagnostic capabilities and hence, prognosis for dogs with HSA, two types of biomarker are needed: a point-of-care diagnostic biomarker and a prognostic biomarker—preferentially based on samples obtained with minimally invasive methods. In this study, we applied a lectin magnetic bead array-coupled tandem mass spectrometry (LeMBA-MS/MS) workflow through discovery and validation phases to discover serum glycoprotein biomarker candidates for canine HSA. By this approach, we found that Datura stramonium (DSA), wheat germ agglutinin (WGA), Sambucus nigra (SNA), and Pisum sativum (PSA) lectins captured the highest number of validated candidate glycoproteins. Secondly, we independently validated serum LeMBA-MS/MS results by demonstrating the in situ relationship of lectin-binding with tumor cells. Using lectin-histochemistry and immunohistochemistry (IHC) for key proteins on tissues with HSA and semi-quantitation of the signals, we demonstrate that a combination of DSA histochemistry and IHC for complement C7 greatly increases the prospect of a more specific diagnosis of canine HSA. Full article

Tick-borne diseases are a major threat to both humans and their pets; therefore, it is important to evaluate the prevalence of pathogens carried by ticks on companion animals. In this study, attached and unattached Ixodid ticks were removed from companion animals by a veterinary practice in Hall County, Georgia. DNA was extracted from unengorged adult ticks and each was screened for the presence of Rickettsia spp. by polymerase chain reaction (PCR) and sequenced to determine the species present. Two hundred and four adult hard-bodied ticks were identified to species and Rickettsia spp. were found in 19.6% (n = 38) of the 194 analyzed DNA extracts. Rickettsia montanensis was found in Dermacentor variablis (14.7%; n = 25), Amblyomma maculatum (33.3%; n = 2), and Rhipicephalus sanguineus s.l. ticks (25%; n = 4). One Amblyomma americanum tick contained Rickettsia amblyommatis, while Rickettsia felis was found in one Dermacentor variablis tick, serving as the first report of Rickettsia felis in a tick in this region and within this tick vector. This study suggests that there is a risk of companion animals contracting a species of Rickettsia from a tick bite in northeastern Georgia, indicating a need for more investigation and highlighting the importance of tick prevention on pets. Full article

Ticks are one of the most important arthropod vectors and reservoirs as they harbor a wide variety of viruses, bacteria, fungi, protozoa, and nematodes, which can cause diseases in human and livestock. Due to their impact on human, livestock, and wild animal health, increased knowledge of ticks is needed. So far, the published data on the molecular diversity between hard ticks species collected in Tanzania is scarce. The objective of this study was to determine the genetic diversity between hard tick species collected in the wildlife-livestock interface ecosystem at Mikumi National Park, Tanzania using the mitochondrion 16S rRNA gene sequences. Adult ticks were collected from cattle (632 ticks), goats (187 ticks), and environment (28 ticks) in the wards which lie at the border of Mikumi National Park. Morphological identification of ticks was performed to genus level. To identify ticks to species level, molecular analysis based on mitochondrion 16S rRNA gene was performed. Ticks representing the two genera (Hyalomma and Rhipicephalus) were identified using morphological characters. Six species were confirmed based on mitochondrion 16S rRNA gene, including Rhipicephalus microplus, Rhipicephalus evertsi, Hyalomma rufipes, Hyalomma truncatum, Hyalomma marginatum, and Hhyalomma turanicum. The presence of different clusters of tick species reflects the possible biological diversity of the hard ticks present in the study region. Further studies are however required to quantify species of hard ticks present in the study region and the country in general over a larger scale. Full article