Vet. Sci., Volume 8, Issue 10 (October 2021) – 42 articles

Canine intestinal lymphangiectasia (IL) is a condition characterized by variably severe gastrointestinal signs, frequently associated with laboratory abnormalities; the research for markers allowing a better understanding of the severity degree and/or obtaining an early diagnosis and/or monitoring is continuously progressing. In the present study, we investigated possible new diagnostic/follow-up markers in IL dogs, namely, serum C-reactive protein, serum bacterial lipopolysaccharide, serum cleaved cytokeratin 18, serum citrulline, and zonulin (in both serum and feces). A fecal proteomic study looking for possible confirmation and/or new marker candidates was also performed. All markers in both substrates, with the exception of serum citrulline, significantly differed between diseased and control dogs. Fecal proteomics allowed the retrieval of three proteins in IL dogs (Fc fragment of IgG-binding protein; transthyretin; proproteinase E) that were not previously found in clinically healthy subjects. Although further studies are needed, C-reactive protein, bacterial lipopolysaccharide, cleaved cytokeratin 18, and zonulin (in both serum and feces) resulted as promising markers for canine IL; similarly, fecal proteomics represents a road worthy of being pursued in the search for candidate biomarkers. Full article

The objective of this study was to evaluate the clinical outcomes and complications of a microendoscopic laminectomy and discectomy (MED) for acute thoracolumbar intervertebral disc extrusions in dogs. Eleven client-owned dogs with acute thoracolumbar intervertebral disc extrusions were included in this retrospective case-series. Dogs were diagnosed with acute thoracolumbar intervertebral disc extrusions using computed tomography (CT) and magnetic resonance imaging (MRI). MED was performed with an integrated endoscopic system to the affected intervertebral disc. Surgery time, intra-operative complications, causes of conversion to microscopic surgery if necessary, post-operative complications, and neurological status on presentation at discharge, as well as any further evaluations in hospital, and long-term concerns via owner contact, were recorded. Post-operative CT images were obtained to compare the extent of laminectomy performed to the planned region of laminectomy. The fully endoscopic procedure was completed in eight dogs without major complications. Three cases were converted to an open surgery due to difficulty removing extruded disc material and controlling hemorrhage. The clinical outcome was good in all cases and equivalent to previously reported prognoses after open surgery. MED is an effective and safe alternative to conventional open procedures in dogs with acute thoracolumbar intervertebral disc extrusion. Full article

Anthelmintics are frequently administered to animals to limit fecal egg elimination, so that wild animals in captive breeding programs are treated to maintain a proper health condition. This is effective from a health management perspective, but on the other hand, it could prevent captive animals from developing an effective immunity against parasites that they might encounter when reintroduced into their original geographic areas. The aim of this study was to describe the dynamics of parasite infections in captive Cuvier’s gazelles (Gazella cuvieri) not treated with anthelmintics for two years and to evaluate the factors related to their fecal egg shedding. Fifteen one-year-old males were enclosed together and captured monthly to collect feces directly from the rectum. Fecal egg counts were performed, and eggs were classified as strongylid-like, Nematodirus sp., or Trichuris sp. Fecal egg shedding for the three groups of parasites did not vary significantly over the duration of the study. Only precipitation affected the egg-shedding pattern of all parasites, while inbreeding was positively associated with the number of strongylid-like parasites. These findings suggest an equilibrium between hosts and parasites in absence of treatment during the study. The anthelmintic treatment as a systematic prophylaxis method in captive animals should be avoided and replaced by systematic coprological and clinical vigilance, as well as targeted treatment in the case of a significant rise of fecal egg counts. Full article

Infectious bronchitis (IB) of chickens is a highly contagious disease characterized by damage of the respiratory system and reproductive organs in young animals caused by a virus of the genus Gamma coronavirus. The condition of the respiratory system caused by the IB virus in chickens has many similarities with the pathology of the respiratory system caused by SARS-CoV-2 in humans. The effectiveness of virucidal drugs (Argovit, Triviron, Ecocid, and lauric acid monoglyceride) was tested on chickens inoculated with a tenfold dose of a vaccine strain based on the attenuated virus H120 against IB of chickens. On the 6th day after inoculation, inflammatory changes in the intestines, lungs, and thymus were observed in the control group. The experimental groups were characterized by less pronounced inflammatory reactions and a lower proportion of thymus and lung probes containing genomic IB virus RNA. Since the virucidal activity of four orally administrated formulations was possible only in the intestine, the experimental data indirectly confirmed the hypothesis of the possibility of the predominant accumulation of coronaviruses in the intestine and subsequent lung damage due to the hematogenous redistribution of viral particles and IBV antigens. It was suggested that other coronaviruses including SARS-CoV-2 can implement a similar mechanism. Full article

This study aimed to compare the in vitro and in vivo retention, bacterial adhesion, and biofilm formation between anionic and zwitterionic bandage contact lenses (BCLs) in healthy canines. BCL retention and tolerance were evaluated in 10 healthy canines via a single-masked, crossover study for 7 days. To compare in vitro bacterial adhesion and biofilm formation, four Staphylococcus strains were incubated with the BCLs at 37 °C for 2 or 24 h, and the bacterial colony forming units (CFUs) adhering to the BCLs were counted. Next, to compare in vivo bacterial adhesion, the CFUs of bacteria adhering to the BCLs worn by canines for 24 h were counted. Anionic lenses significantly retained and reduced in vitro bacterial adhesion than in the zwitterionic lenses. However, the amount of in vitro biofilm formation was more likely to be higher on anionic lenses than on zwitterionic lenses. In vivo bacterial adhesion was not significantly different between the two types of BCLs. Nevertheless, both BCLs were well-tolerated by the canines; thus, their short-term use in dogs can be recommended as safe. Full article

Resistance to anthelmintic drugs in gastrointestinal nematodes (GIN) of sheep is of high concern for livestock production worldwide. In Calabria (southern Italy), many plants have been used in ethnoveterinary medicine for parasite control in small ruminants. Here, we present an in vivo evaluation of anthelmintic efficacy of three plant extracts. The first was based on bark and leaves of Salix caprea, the second and the third were based on the whole plant Artemisia campestris and whole fruit (seeds and peel) of Punica granatum, respectively. Anthelmintic efficacy was evaluated according to the fecal egg count reduction test (FECRT) performed with the FLOTAC technique. The results showed a significant anthelmintic effect of Punica granatum macerate (50%), a low effectiveness of the Artemisia campestris macerate (20%), and a complete ineffectiveness of Salix caprea macerate (0.1%). With these outcomes, we report a P. granatum-based remedy reducing 50% GIN egg output. This result was obtained without using any synthetic drug, paving the way for the employment of green veterinary pharmacology (GVP) as a complementary and sustainable method to reduce the use of chemicals and to counteract anthelmintic resistance. Full article

Background: Canine leishmaniasis (CanL) is caused by Leishmania infantum (L. infantum) that is transmitted by sand fly vectors with dogs acting as the main reservoir. Methods: The present study aimed to determine the seroprevalence of CanL in dogs from Egypt and assessed the associated risk factors. The study was conducted from 2019 to 2020 in five governorates situated in Northern Egypt. Serum samples from 450 asymptomatic dogs were serologically examined by use of enzyme-linked immunosorbent assay (ELISA). Results: Overall, the seroprevalence rate of CanL was 21.3% and the highest rates were observed in Cairo and Giza governorates. The univariable analysis revealed that the seropositivity of CanL was strongly related to the dogs’ ages, length of hair, absence of veterinary care or application of insecticides, and the type of floor of their shelters. The risk factors that were found to be associated with CanL in exposed dogs were: age group 2–4 years old (OR = 12, 95% CI: 1.6–92.3); short hair (OR = 2.07, 95% CI: 1.2–3.6); absence of veterinary care (OR = 2.7, 95% CI: 1.3–5.8); no application of insecticides (OR = 3.09, 95% CI: 1.5–6.5) and their residence in a shelter with an earthen floor (OR = 1.42, 95% CI: 0.7–2.9). Conclusions: Based on the present results, CanL is present in Egyptian dogs and this increases the possibility of transmission by sand fly to humans with whom they have contact. Consequently, an efficient monitoring programme and effective control measures are important to reduce the risk of infection. Full article

Betaarterivirus suid 1 and 2 are the causative agents of porcine reproductive and respiratory syndrome (PRRS), which is one of the most significant diseases of the swine industry, causing significant economic losses in the main pig producing countries. Here, we report the development of a novel, RNA-based in situ hybridization technique (RNAscope) to detect PRRS virus (PRRSV) RNA in lung tissues of experimentally infected animals. The technique was applied to lung tissues of 20 piglets, which had been inoculated with a wild-type, highly pathogenic PRRSV-1 strain. To determine the RNAscope’s applicability as a semi-quantitative method, we analysed the association between the proportion of the virus-infected cells measured with an image analysis software (QuPath) and the outcome of the real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR) tests performed in parallel. The results of the quantitative approach of these two molecular biological methods show significant association (pseudo R² = 0.3894, p = 0.004). This is the first time RNAscope assay has been implemented for the detection of PRRSV-1 in experimental animals. Full article

(1) Background: Accounting for the well-being of equine partners is a responsibility of those engaged in Equine-Assisted Services (EAS). Researchers took heed of this call to action by developing an innovative way to collect data to assess the physiological indicators of stress in equine participants. The collection of saliva is considered to be a minimally invasive method of data collection and is typically performed using a cotton swab; however, in equines, the introduction of a foreign object may induce stress; (2) Methods: Researchers used a modified bit to collect pooled saliva in an effort to further reduce stress during the saliva collection process. Additionally, the collection of pooled saliva, via the bit, increases the opportunity to consider additional analyses, such as oxytocin, which is more reliable in pooled saliva than site-specific saliva captured with a swab; (3) Results: A data analysis demonstrated that ample saliva was captured using the modified bit. Observational data supported that the horses demonstrated fewer physical stress signals to the bit than to the swab. Thus, the modified bit is a feasible and valid method for equine salivary sample collection; (4) Conclusions: The results suggest that the modified bit provides a viable method to collect equine saliva and supports national calls to prioritize animal welfare analysis, specifically for horses used within EAS. Future research should enhance methodological rigor, including in the process and timing, thereby contributing to the bit’s validation. Full article

Bones play an important role in maintaining the level of calcium in blood. They provide support for soft tissues and hematopoiesis and undergo continuous renewal throughout life. In addition, vitamin D is involved in regulating bone and calcium homeostasis. Galectin-3 (Gal-3) is a β-galactoside-binding protein that can regulate bone cell differentiation and function. Here, we aimed to study the regulatory effects of Gal-3 on vitamin-D-regulated osteoclastogenesis and bone resorption in chicken. Gal-3 expression in bone marrow stromal cells (BMSCs) from 18-day-old chicken embryos was inhibited or overexpressed. BMSCs were then co-cultured with bone marrow monocytes/macrophages (BMMs) with or without addition of 1α,25(OH)₂D₃. The results showed that 1α,25(OH)₂D₃ upregulated the expression of Gal-3 mRNA and receptor activator of nuclear-factor κB ligand (RANKL) expression in BMSCs and promoted osteoclastogenesis, as shown by the upregulated expression of osteoclast (OC) markers (CtsK, CAII, MMP-9, and TRAP) and increased bone resorption, a method for measuring the bone resorption area in vitro. Knockdown of Gal-3 by small-interfering RNA (siRNA) in BMSCs downregulated the expression of RANKL mRNA and attenuated the effects of 1α,25(OH)₂D₃ on osteoclastogenesis and bone resorption. Conversely, overexpression of Gal-3 in BMSCs enhanced the effects of osteoclastogenesis and bone resorption by increasing the expression of RANKL mRNA. These results demonstrated that Gal-3 mediates the differentiation and bone resorption of osteoclasts regulated by 1α,25(OH)₂D₃. Full article

Maternal diet during early gestation affects offspring phenotype, but it is unclear whether maternal diet during late gestation influences piglet metabolism. We evaluated the impact of two dietary protein levels in sow late gestation diet and piglet nursery diet on piglet metabolism. Diets met or exceeded the crude protein and amino acid requirements. Sows received either 12% (Lower, L) or 17% (Higher, H) crude protein (CP) during the last five weeks of gestation, and piglets received 16.5% (L) or 21% (H) CP from weaning at age 3.5 weeks. This resulted in a 2 × 2 factorial design with four sow/piglet diet treatment groups: HH and LL (match), HL and LH (mismatch). Piglet hepatic tissues were sampled and differentially expressed genes (DEGs) were determined by RNA sequencing. At age 4.5 weeks, 25 genes were downregulated and 22 genes were upregulated in the mismatch compared to match groups. Several genes involved in catabolic pathways were upregulated in the mismatch compared to match groups, as were genes involved in lipid metabolism and inflammation. The results show a distinct interaction effect between maternal and nursery diets, implying that sow late gestation diet could be used to optimize piglet metabolism. Full article

Blood samples were obtained from 16 high-risk heifers; eight were newly arrived from a 40 h road trip (0 days post-arrival (DPA)), whereas the other eight heifers had been in the feedlot at 25 DPA. Both groups were transported from the southeast tropical region of Mexico to a feedlot in the northeast and were sampled on the same day. The complete blood count, blood chemistry, and cytokine gene expression were analyzed. Gene expression was analyzed using specific primers to amplify and quantify the cDNA reverse transcribed from the mRNA transcripts for tumor necrosis factor (TNF)-α, interferon (IFN)-γ, and interleukin (IL)-2. Higher values for hematocrit (p = 0.029), hemoglobin (p = 0.002), eosinophils (0.029), albumin (p = 0.014), alanine aminotransferase (p = 0.004), bilirubin (p = 0.003), cholesterol (p = 0.014), and cortisol (p = 0.051) were observed in the 0 DPA group than the 25 DPA group. In the electrophoresis of TNF-α amplification products, two non-specific bands were observed in the 0 DPA group. These bands were sequenced, and BLAST analysis suggested that they corresponded to bovine lymphotoxin and have not been reported previously related to stress. The TNF-α expression level was higher (p = 0.001) in the 25 DPA group than the 0 DPA group according to the semi-quantitative expression analysis. This may indicate a persistent inflammatory process that could be related to trauma and disease, which can negatively impact their subsequent health and growth performance. In conclusion, homeostatic disruption was apparent in the 0 DPA heifers, which showed higher cortisol and reductions in TNF-α levels and stress-induced bovine lymphotoxin (SIBL) co-expression. Full article

Cardiac arrest (CA) causes severe spinal cord injury and evokes spinal cord disorders including paraplegia. It has been reported that risperidone, an antipsychotic drug, effectively protects neuronal cell death from transient ischemia injury in gerbil brains. However, until now, studies on the effects of risperidone on spinal cord injury after asphyxial CA (ACA) and cardiopulmonary resuscitation (CPR) are not sufficient. Therefore, this study investigated the effect of risperidone on hind limb motor deficits and neuronal damage/death in the lumbar part of the spinal cord following ACA in rats. Mortality, severe motor deficits in the hind limbs, and the damage/death (loss) of motor neurons located in the anterior horn were observed two days after ACA/CPR. These symptoms were significantly alleviated by risperidone (an atypical antipsychotic) treatment after ACA. In vehicle-treated rats, the immunoreactivities of tumor necrosis factor-alpha (TNF-α) and interleukin 1-beta (IL-1β), as pro-inflammatory cytokines, were increased, and the immunoreactivities of IL-4 and IL-13, as anti-inflammatory cytokines, were reduced with time after ACA/CPR. In contrast, in risperidone-treated rats, the immunoreactivity of the pro-inflammatory cytokines was significantly decreased, and the anti-inflammatory cytokines were enhanced compared to vehicle-treated rats. In brief, risperidone treatment after ACA/CPR in rats significantly improved the survival rate and attenuated paralysis, the damage/death (loss) of motor neurons, and inflammation in the lumbar anterior horn. Thus, risperidone might be a therapeutic agent for paraplegia by attenuation of the damage/death (loss) of spinal motor neurons and neuroinflammation after ACA/CPR. Full article

Immunohistochemistry (IHC) is a widely used technique in diagnostic pathology, but the simultaneous analysis of more than one antibody at a time with different chromogens is rather complex, time-consuming, and quite expensive. In order to facilitate the identification of mast cells (MCs) during immunohistochemical analysis of membrane and/or nuclear markers, we propose a new staining method that includes the association of IHC and toluidine blue as a counterstain. To achieve this goal, we tested c-kit, Ki67, and cannabinoid receptor 2 on several cases of cutaneous canine mast cell tumors (MCTs), cutaneous mastocytosis, and atopic dermatitis. The results obtained show how this double staining technique, although limited to non-cytoplasmic markers and of little use in poorly differentiated MCTs in which MC metachromasia is hard to see, can be used during the evaluation of nuclear and/or membranous immunohistochemical markers in all canine cutaneous disorders, especially if characterized by the presence of a low number of MCs. It can help to evaluate those MCTs in which neoplastic MCs must be clearly distinguished from inflammatory cells that can infiltrate the tumor itself, in facilitating the calculation of the Ki67 index. Moreover, it can be used to study the expression of new markers in both animal and human tissues containing MCs and in MC disorders. Full article

Globally, the use of ethnoveterinary medicine as remedies for animal health among different ethnic groups justify the need for a systematic exploration to enhance their potential. In addition, the increasing popularity and utilisation of woody plants remain common in traditional medicine, which may be attributed to their inherent benefits. The current review was aimed at analysing ethnoveterinary surveys, biological activities, and secondary metabolites/phytochemical profiles of the woody plants of South Africa. Eligible literature (period: 2000 to 2020) were retrieved from different databases such as Google Scholar, PubMed, Sabinet, and Science Direct. Based on the inclusion and exclusion criteria, 20 ethnoveterinary surveys were eligible and were subjected to further analysis. We identified 104 woody plant species from 44 plant families that are used in the treatment of different diseases in animals, particularly cattle (70%) and goats (20%). The most mentioned (with six citations) woody plants were Terminalia sericea Burch. ex DC and Ziziphus mucronata Willd., which were followed by plants with five (Cussonia spicata Thunb., Pterocarpus angolensis DC and Vachellia karroo (Hayne) Banfi & Galasso) or four (Acokanthera oppositifolia (Lam.) Codd, Cassia abbreviata Oliv., and Strychnos henningsii Gilg) individual mentions. The most dominant families were Fabaceae (19%), Apocynaceae (5.8%), Rubiaceae (5.8%), Anacardiaceae (4.8%), Combretaceae (4.8%), Euphorbiaceae (4.8%), Malvaceae (4.8%), Rhamnaceae (4.8%), and Celastraceae (3.8%). Bark (33%), leaves (29%), and roots (19%) were the plant parts dominantly used to prepare remedies for ethnoveterinary medicine. An estimated 20% of woody plants have been screened for antimicrobial, anthelmintic, antioxidant, and cytotoxicity effects. Phytochemical profiles established a rich pool of valuable secondary metabolites (phenolic, flavonoids and condensed tannins) that may be responsible for the exerted biological activities. Overall, the significant portion of woody plants lacking empirical evidence on their biological effects indicates a major knowledge gap that requires more research efforts. Full article

A two-year-and-eleven-month-old male Shikoku Inu was referred for evaluation of progressive gait abnormality that had begun three months prior. Neurological examination revealed ventral flexion of the neck, a wide-based stance in the hindlimb, wide excursions of the head from side to side, tremor in all four limbs, hypermetria in all four limbs, proprioceptive deficits in all four limbs, reduced patellar reflex in both hindlimbs, and postural vertical nystagmus. Later, behavioral and cognitive dysfunction, ataxia, and visual deficits slowly progressed. Magnetic resonance imaging revealed symmetrical progressive atrophy of the whole brain and cervical spinal cord. Bilateral retinal degeneration was observed, and both flush and flicker electroretinograms were bilaterally non-recordable at the age of five years and eight months, and the dog was euthanized. Histopathologically, faint-to-moderate deposition of light-brown pigments was frequently observed in the cytoplasm of neurons throughout the cerebrum, cerebellum, and nuclei of the brainstem. The pigments were positive for Luxol fast blue, periodic acid–Schiff, and Sudan black B, and exhibited autofluorescence. Electron microscopic examination revealed the accumulation of membranous material deposition in the neuronal cytoplasm. Small foci of pigment-containing macrophages were frequently observed around the capillary vessels. Based on these clinical and pathological findings, the animal was diagnosed with adult-onset neuronal ceroid lipofuscinosis. Full article

During the past decade, the field of human–animal interaction(s) research has been characterized by a significant increase in scientific findings. These data have contributed to our current understanding of how humans may benefit from contact with animals. However, the animal experience of these interactions is still an under-researched area. This paper addresses the welfare of dogs who participate in animal-assisted interventions (AAIs) to improve health in human recipients. This paper builds on previous work by Glenk (2017) and provides an updated review of the literature on therapy dog welfare published from 2017–2021. New advances in scientific methodology, such as the determination of salivary oxytocin, breath rate and tympanic membrane temperature, are analyzed regarding their value and limitations for research in AAIs. Moreover, welfare-related social and environmental factors (e.g., freedom of choice, exploration of novel environments, inequity aversion, individual development, working experience, relationship with handler and handler skills) that profoundly influence dog perception and well-being are reviewed and discussed. Accounting for the globally increasing interest and the number of dogs utilized in AAIs, safeguarding therapy dog well-being, and identifying situations, circumstances and protocols that may challenge animal welfare remains an emerging and crucial area of scientific effort. Full article

Symphyseal distraction osteotomy (SDO) with a polymethyl methacrylate (PMMA) spacer is an effective surgical treatment for cats with pelvic stenosis. This study reports the successful treatment of urethral obstruction due to ischiocavernosus muscle (IM) tension after SDO with a PMMA spacer. A 2-year-old castrated male Korean domestic shorthair feline had megacolon and pelvic canal stenosis. The ratio of the maximal diameter of the colon to the L5 length and the pelvic canal diameter ratio were 1.6 and 0.45, respectively. Pelvic SDO was performed with a PMMA spacer, leading to pelvic canal enlargement (pelvic canal diameter ratio: 0.73). Two days after surgery, dysuria was identified immediately after removing the preoperatively placed urinary catheter. Complete blood counts and serum biochemical profiles were within the reference intervals, and a positive contrast retrograde urethrogram confirmed urethral obstruction at the level of the membranous–pelvic urethra region. Increased tension of the IM leading to a narrowed urethra was suspected as the cause of urethral obstruction. After IM release at the level of origin on the ischium, the patient had an uneventful recovery with spontaneous urination. Muscle release resulted in excellent functional restoration, with no intraoperative or postoperative complications reported during the 12-month long-term follow-up. Therefore, SDO with IM release could be a feasible therapeutic option for severe pelvic stenosis without complications, such as urethral obstruction, in cats. Full article

A two-year-old ball python with a submandibular mass was evaluated. Fine needle aspiration resulted in debris containing purulent materials and bacterial cells on cytology. Radiography demonstrated multi-focal radiopaque lesions in the mass, which were suspected to be mineralization; there was an absence of mandibular invasion or lung involvement. Gross examination of the surgically excised mass revealed a multi-nodular, well-circumscribed lesion with purulent material. The postoperative recovery was uneventful. The histopathological examination followed by immunohistochemistry analysis gave a diagnosis of leiomyosarcoma. As tumors containing purulent materials can be confused with an abscess, diagnostic confirmation with various diagnostical tools should be considered. Full article

African swine fever (ASF) is a devastating viral disease in pigs and is therefore economically important for the swine industry. ASF is characterized by a short incubation period and immediate death, making the early identification of ASF-infected pigs essential. This pilot-scale study evaluates whether the infrared thermography (IRT) technique can be used as a diagnostic tool to detect changes in skin temperature (T_sk) during the early stages of disease development in experimentally ASF-infected pigs. Clinical symptoms and rectal temperatures (T_core) were recorded daily, and IRT readings during the experimental ASF infection were analyzed. All infected pigs died at 5–8 days post inoculation (dpi), and the incubation period was approximately 4 dpi. The average T_core increased from 0 dpi (38.9 ± 0.3 °C) to 7 dpi (41.0 ± 0.5 °C) and decreased by 8 dpi (39.8 ± 0 °C). The maximum T_sk of ASF-infected pigs increased from 2 (35.0 °C) to 3 dpi (38.5 °C). The mean maximum T_sk observed from three regions on the skin (ear, inguinal, and neck) significantly increased from 2 to 3 dpi. This study presents a non-contact method for the early detection of ASF in infected pigs using thermal imaging at 3 days after ASF infection. Full article

Development of in vitro culture and completion of genome sequencing of several Babesia parasites promoted the efforts to establish transfection systems for these parasites to dissect the gene functions. It has been more than a decade since the establishment of first transfection for Babesia bovis, the causative agent of bovine babesiosis. However, the number of genes that were targeted by genetic tools in Babesia parasites is limited. This is partially due to the low efficiencies of these methods. The recent adaptation of CRISPR/Cas9 for genome editing of Babesia bovis can accelerate the efforts for dissecting this parasite’s genome and extend the knowledge on biological aspects of erythrocytic and tick stages of Babesia. Additionally, glmS ribozyme as a conditional knockdown system is available that could be used for the characterization of essential genes. The development of high throughput genetic tools is needed to dissect the function of multigene families, targeting several genes in a specific pathway, and finally genome-wide identification of essential genes to find novel drug targets. In this review, we summarized the current tools that are available for Babesia and the genes that are being targeted by these tools. This may draw a perspective for the future development of genetic tools and pave the way for the identification of novel drugs or vaccine targets. Full article

Hyperlipidemia is a risk factor for nonalcoholic fatty liver disease (NAFLD) in humans. However, the association between serum lipids and canine chronic hepatitis remains unknown. In this study, serum lipids, hepatic profiles, and hepatic ultrasound scores of healthy dogs and dogs with chronic hepatitis were evaluated. Serum triglyceride and cholesterol concentrations were significantly higher (p < 0.01) in dogs with chronic hepatitis. There were 62.2% of dogs with chronic hepatitis accompanied by hypertriglyceridemia, hypercholesterolemia, or both. Positive correlations were observed between serum ALT and cholesterol (r = 0.8287, p < 0.01), serum ALP and cholesterol (r = 0.8436, p < 0.01), serum GGT and cholesterol (r = 0.5640, p < 0.01), serum bile acid and cholesterol (r = 0.3310, p < 0.01) and serum ALP and triglycerides (r = 0.2582, p < 0.05). No significant differences were found between ultrasound scores of diseased dogs with and without hypertriglyceridemia and diseased dogs with and without hypercholesterolemia. Canine chronic hepatitis is associated with hyperlipidemia. A significant positive association was identified between hyperlipidemia, especially hypercholesterolemia, liver enzymes, and bile acid concentration in dogs suffering from chronic hepatitis. The underlying mechanisms connecting hyperlipidemia and canine chronic hepatitis remain elusive. Full article

The treatment of choice for feline hyperthyroidism is the administration of the antithyroid drug methimazole. Both the endocrinopathy and the drug adverse reactions (e.g., hepatotoxicosis, gastrointestinal disorders, and renal injury) are partly due to oxidative stress and redox unbalance. This study investigated the free radical production and the impairment of the antioxidant barrier induced by methimazole in an in vitro model of feline renal epithelium. The protective effects of quercetin and resveratrol were also explored. CRFK cells were incubated with a methimazole concentration equivalent to the maximum plasma levels in orally treated cats (4 µM), in the presence or absence of either one of the two selected antioxidants at different time-points (up to 72 h). Cell viability, ROS production, GSH levels, and mRNA expression of antioxidant enzymes (i.e., CAT, SOD, GPx, and GST) were assessed. Methimazole impaired cell viability and increased ROS levels in a time-dependent manner. Similarly, GSH content and CAT, SOD, and GPx3 expression were higher compared with control cells. Such effects were significantly counteracted by quercetin. These results provide new insights about the mechanisms underlying the methimazole-related side effects frequently observed in hyperthyroid cats. They also support the use of quercetin in the management of feline hyperthyroidism. Full article

The objectives of this study were to investigate temperature distribution at the sheep hoof and evaluate the reliability and diagnostic performance of infrared thermography (IRT) for the detection of footrot and white line disease (WLD) lesions in intensively reared dairy sheep. Hoof lesions were clinically assessed, and IRT was used to measure temperature distribution on hoof superficial tissue in 600 multiparous ewes. Binary regression models were developed and validated, and receiver operating characteristic curves were estimated to assess the predictive value and diagnostic performance of IRT for the detection of hoof lesions. The most sensitive prediction model for the detection of IFR was based on the difference between ambient and hoof heel temperature (sensitivity: 83.3%, specificity: 47.8%, and threshold value: 6.5 °C), whereas the most specific prediction model was based on the difference between ambient and coronary band temperature (sensitivity: 51.9%, specificity: 79.7%, and threshold value: 11.3 °C). In the case of WLD, the diagnostic performance of IRT had limited predictive value. IRT could be a useful tool for hoof health screening in dairy sheep. However, it must be cautiously adapted in cases where environmental, operating, and operator variables are not effectively controlled. Full article

Pesticides can induce changes in behavior and reduce the survival chance of aquatic organisms. In this study, the toxic effects of glyphosate suspension (Glyphosate Aria 41% SL, Tehran Iran) on behavior and tissues of common carp (Cyprinus carpio) were assessed. For this purpose, a 96 h LC₅₀ of glyphosate suspension (68.788 mL·L⁻¹) was used in the toxicity test. All individuals were divided into control and treatment groups with four replicates. Exposure operations were performed under two conditions: increasing concentration of suspension from 0 to 68.788 mL·L⁻¹; then, decreasing to the first level. The swimming pattern was recorded by digital cameras during the test and tissue samples were collected at the end of the test. There were significant differences between the swimming pattern of treated individuals and control ones during both steps. The sublethal concentration of glyphosate led to hypertrophy, hyperplasia and hyperemia in the gill of fish. However, changes were obvious only after sampling. The exposed fish also displayed clinical signs such as darkening of the skin and increasing movement of the operculum. Moreover, glyphosate suspension affected swimming patterns of fish suggest that the swimming behavior test can indicate the potential toxicity of environmental pollutants and be used as a noninvasive, useful method for managing environmental changes and assessing fish health conditions by video monitoring. Full article

To reduce the circulation path of the output current of traditional electroacupuncture (TEA) process in the body, a simple single-acupoint electroacupuncture (SEA) frame was designed and the acupuncture effect of SEA was evaluated through Hou-san-li (ST-36) and Qian-san-li (LI-10) acupoints. Forty-two healthy New Zealand rabbits were randomly divided into seven groups and underwent acupuncture for 20 min in an awake state. Blood samples aseptically collected from the ear vein 3 h before acupuncture and 0, 3, 6, 9, 12 and 24 h after acupuncture were used for the detection of aspartate aminotransferase (AST), alanine aminotransferase (ALT), creatine kinase MB (CK-MB) and motilin (MTL) in serum. The simple SEA frame was developed successfully, and the acupuncture results showed that the serum AST and ALT levels were significantly higher at 3 h after TEA with high frequency (p < 0.01) compared with the control group. Regarding serum CK-MB levels, no significant differences were found after SEA or TEA stimulation (p > 0.05). Serum MTL levels were significantly increased at 0 h after SEA and TEA (p < 0.05), but there were no significant differences at other time points after SEA and TEA treatment (p > 0.05). SEA not only maintains the effect of TEA but also shortens the circulation loop of the electroacupuncture (EA) current in the body, which effectively avoids body injury. Full article

Teladorsagia circumcincta is the most important gastrointestinal parasite in the livestock industry in temperate regions around the world, causing great economic losses. The infective third-stage larvae (L3) of Teladorsagia circumcincta secrete a large number of excretory-secretory (E/S) molecules, some of which are likely to play critical roles in modulating the host immune response. One of the most abundant E/S molecules is a protein termed Tci-gal-1, which has similarity to mammalian galectins. Galectins are a family of carbohydrate-binding molecules, with characteristic domain organisation and affinity for β-galactosids that mediate a variety of important cellular functions including inflammation and immune responses. To understand the role of Tci-gal-1 at the host–parasite interface, we used a proteomics pull-down approach to identify Tc-gal-1 interacting proteins from sheep abomasal scrapes and whole tissue. A total of 135 unique proteins were identified from whole abomasal tissue samples, while 89 proteins were isolated from abomasal scrape samples. Of these proteins, 63 were present in both samples. Many of the host proteins identified, such as trefoil factors and mucin-like proteins, play critical roles in the host response. The identification of Tci-gal-1 binding partners provides new insights on host–parasite interactions and could lead to the development of new control strategies. Full article

Leishmaniasis remains one of the major neglected tropical diseases. The epidemiological profile of the disease comprises a wide range of hosts, including dogs and cats. Despite several studies about feline Leishmaniosis, the role of cats in disease epidemiology and its clinical impact is still debated. The present study raises awareness about the impact of leishmaniasis in cats from an endemic region in of Northwestern Italy (Liguria). A total number of 250 serum and 282 blood samples were collected from cats, then assessed for Leishmania infantum (L. infantum) serologically using western blot (WB) and molecularly using polymerase chain reaction (PCR). We also tested the association of Leishmania infection with some infectious agents like haemotropic Mycoplasma, Feline immunodeficiency virus (FIV) and Feline leukemia virus (FeLV) together with the hematobiochemical status of the examined animals. Interestingly, all tested animals were asymptomatic and out of 250 examined serum samples, 33 (13.20%) samples (confidence interval (CI) 95% 9.56–17.96%) were positive at WB for L. infantum, whereas of the 282 blood samples, 80 (28.36%) returned a positive PCR (CI 95% 23.43–33.89%). Furthermore, there was a statistical association between PCR positivity for L. infantum and some hematological parameters besides FIV infection as well as a direct significant correlation between Mycoplasma infection and WB positivity. Taken together, the present findings report high prevalence of L. infantum among cats, which reinforces the significance of such positive asymptomatic animals and confirms the very low humoral response in this species. In addition, the laboratory values provide evidence that infection by the parasite is linked to alteration of some hematological parameters and is correlated to some infectious agents. These data are of interest and suggest future research for accurate diagnosis of such zoonosis. Full article

The aim of this study was to report the results of autologous bone marrow mononuclear cell (BMMC) transplantation as a minimally invasive treatment for grade 2 UAP in dogs. This was an observational case series on six German shepherd dogs affected by grade 2 UAP as defined according to their clinical condition as well as radiographic and CT findings. Bone marrow was collected from the iliac crest and the mononuclear fraction was separated with density gradient centrifugation. Cells were suspended in fibrin glue before BMMC administration and implanted via transcutaneous injection under IB or CT guidance, using a spinal needle directly inserted into the ossification centre between the anconeal process and the olecranon. Clinical and radiographic follow-up was performed for up to 6 months. Microradiographic assessment was performed on one dog that died of other causes. A progressive reduction of pain within 3 weeks after BMMC administration was observed in all dogs, with gradually increased weight bearing on the affected limb. Radiographic and CT follow-up revealed the progressive fusion of the ossification centre at 90 days without any signs of secondary OA. The examination of microradiographs showed newly formed bone tissue in which a residue of calcified cartilage was present at the site of BMMC implantation. On the basis of these results, BMMC therapy for grade 2 UAP may be considered to be an effective and minimally invasive treatment option for dogs. Full article

Platelet-rich fibrin (PRF) is a recent platelet-based biomaterial, poised as an innovative regenerative strategy for the treatment of wounds from different etiologies. PRF is defined as a biodegradable scaffold containing elevated amounts of platelets and leukocytes having the capability to release high concentrations of bioactive structural proteins and acting as a temporal release healing hemoderivative. This study aimed to evaluate the performance of canine-origin PRF, obtained from blood of screened donors, as a regenerative biomaterial suitable for the treatment of critical wounds in felines. Four short-hair felines with naturally occurring wounds were enrolled in this study. Three of the wounds were considered infected. Each PRF treatment was the result of the grafting of newly produced PRFs at the recipient area. The PRF treatment was initially performed two to three times per week, followed by single weekly treatments. The study was finalized when complete wound closure was achieved. No topical antimicrobial/antiseptic treatment was applied. The present research demonstrated that xenogenic PRFs significantly induced healthy vascularized granulation tissue in lesions with soft tissue deficit, also prompting the epithelization at the injured site. No rejection, necrosis, or infection signs were recorded. Additionally, PRF-therapy was revealed to be a biological cost-effective treatment, accelerating the wound healing process. Full article