Vet. Sci., Volume 10, Issue 12 (December 2023) – 46 articles

The utilization of Autologous Conditioned Serum (ACS) for treating osteoarthritis (OA) in horses has seen a notable increase in recent years. In vitro studies have consistently identified ACS as a promising therapy for OA joints, contributing to its growing popularity in OA treatment. Despite this, there is a noticeable absence of systematic reviews focused solely on the clinical data of OA patients treated with ACS, excluding the in vitro perspective. This study aims to address this gap by systematically reviewing the latest literature, concentrating solely on clinical data in in vivo studies to evaluate the efficacy of ACS in OA lesions. All clinical studies involving ACS treatments for horses with OA were included in the assessment. Surprisingly, only six trials met the inclusion criteria for this systematic review. The results indicate that the majority of the considered articles support the use of ACS as a treatment for horses, albeit with a control group provided in only one study. However, the absence of a control group and the exclusion of histological evaluation diminish the validity of the majority of clinical research. While several studies suggest a beneficial effect of ACS on OA horses without significant adverse effects, this systematic review affirms that there is no definitive evidence for its effectiveness. Therefore, further investigation of the efficacy of ACS products as a treatment for OA is warranted, emphasizing the need for more controlled trials. Poorly designed and biased studies, lacking blinding or control and adopting inadequate outcome measures, may favor positive results and, thus, necessitate a more rigorous approach to validate the efficacy of ACS in OA treatment. Full article

Restrictions on the use of antimicrobial compounds have led to a surge of interest in alternative solutions, such as natural, plant-based compounds. In our study, we investigated the efficacy of three feed supplements containing different additives, namely, probiotics (Lactobacillus spp., “Test substance A”), turmeric (Curcuma longa L., “Test substance B”), and fenugreek (Trigonella foenum graecum, “Test substance C”). In the experiment, we tested 180 birds of the Bábolna Tetra-SL laying hybrid breed that were infected with Salmonella enteritidis strains. The birds were randomly divided into six groups: three groups treated with the different additives, a negative control group, a positive control group, and an antibiotic-treated group using enrofloxacin. We examined the maturation and the time course of shedding of Salmonella; at the end of rearing, pathological and histopathological examinations were performed. When Salmonella was isolated from the cloacal swab samples, the enrofloxacin-treated group had a high number of animals shedding Salmonella by day 9, which was like the group treated with test material C. The greatest reduction in Salmonella shedding was observed in the groups treated with test materials A and B. In terms of pathological parameters, villus length and crypt depth were significantly better in the group treated with test material C compared to the positive and negative controls, and when comparing the body weight of the tested animals, the group treated with test material B had a significantly larger absorption surface area compared to the positive control group. Overall, the supplement with test material C proved to be the most effective. In the future, it is worthwhile to investigate the combination of the tested active substances for their possible synergistic effects and to perform a dose-response study to select the optimal dosage. Full article

(1) Background: dental pathologies are the most frequent reason for requesting a CT scan of the head in rabbits and guinea pigs. The study aimed to review head CT exams of both species to identify and characterize lesions secondary to dental disease. (2) Methods: head CT studies of 48 rabbits and 52 guinea pigs with dental pathologies were reviewed. (3) Results: dental abnormalities of mandibular teeth were the most represented, both in rabbits (81.2%) and guinea pigs (98%). The aggressive bone lesion associated with teeth was the more frequently observed bone lesion in rabbits’ mandible and maxilla; in guinea pigs, the more frequent bone lesions were bulging around the roots of the teeth with focal lysis in the maxilla, and without focal lysis in the mandible. In the maxilla, the increased attenuation of nasal cavities (rhinitis) was the most frequently observed abnormality both in rabbits (60%) and guinea pigs (83.3%); the exophthalmos was more represented in rabbits (53.3%). In the mandible, the cavernous space-occupying lesion was more represented both in rabbits (92.3%) and guinea pigs (73.3%). (4) Conclusions: lesions secondary to dental pathologies were often observed both in rabbits and guinea pigs; CT examination has proven to be valuable in detecting secondary alterations in both species. Full article

This study describes the anatomical characteristics of the abdominal and pelvic vascular system of two healthy mature female cats via three-dimensional contrast enhanced computed tomography angiography, non-contrast enhanced magnetic resonance angiography and three-dimensional printing. Volume-rendering computed tomography angiography images were acquired from the ventral aspect using RadiAnt, Amira and OsiriX MD Dicom three-dimensional formats, and three-dimensional printing was obtained and compared with the corresponding computed tomography angiography images. Non-contrast enhanced magnetic resonance angiography was made using the time-of-flight imaging in ventral, oblique and lateral views. In addition, three cadavers with colored latex injection were dissected to facilitate the identification of the vascular structures. Three-dimensional computed tomography angiography showed the main vascular structures, whereas with the time-of-flight blood appeared with a high signal intensity compared with associated abdominal and pelvic tissues. Three-dimensional computed tomography angiography images and time-of-flight sequences provided adequate anatomical details of the main arteries and veins that could be used for future feline anatomical and clinical vascular studies of the abdomen and pelvis. Full article

Lymphoma is the most common haematological malignancy affecting dogs and has a high incidence in the Bullmastiff breed. The aim of this study was to identify risk loci predisposing this breed to the disease. The average age of lymphoma diagnosis in 55 cases was less than 6 years, similar to the median age of 64 cases from our clinical and pathology databases. When fine-scale population structure was explored using NETVIEW, cases were distributed throughout an extended pedigree. When genotyped cases (n = 49) and dogs from the control group (n = 281) were compared in a genome-wide association analysis of lymphoma risk, the most prominent associated regions were detected on CFA13 and CFA33. The top SNPs in a 5.4 Mb region on CFA13 were significant at a chromosome-wide level, and the region was fine-mapped to ~1.2 Mb (CFA13: 25.2–26.4 Mb; CanFam3.1) with four potential functional candidates, including the MYC proto-oncogene bHLH transcription factor (MYC) and a region syntenic with the human and mouse lncRNA Pvt1 oncogene (PVT1). A 380 Kb associated region at CFA33: 7.7–8.1 Mb contained the coding sequence for SUMO specific peptidase7 (SENP7) and NFK inhibitor zeta (NFKBIZ) genes. These genes have annotations related to cancer, amongst others, and both have functional links to MYC regulation. Genomic signatures identified in lymphoma cases suggest that increased risk contributed by the regions identified by GWAS may complement a complex predisposing genetic background. Full article

The most significant ectoparasitic mite of honeybees, Varroa destructor, has a detrimental effect on bee health and honey output. The principal strategy used by the control programs is the application of synthetic acaricides. All of this has resulted in drug resistance, which is now a major worry for beekeeping. As a result, research on alternate products and techniques for mite management is now required. The aim of this study was to determine whether essential oils (EOs) extracted from botanical species of Lamiacae, typical of the Calabria region of Southern Italy, could reduce the population of the mite V. destructor. Among the best-known genera of the Lamiaceae family are oregano, rosemary and thyme, whose EOs were employed in this study. By steam distillation, the EOs were extracted from Origanum vulgare subsp. viridulum (Martrin-Donos) Nyman, Thymus capitatus Hoffmanns. and Link, Thymus longicaulis C.Presl and Salvia rosmarinus Schleid. plant species harvested directly on the Calabrian territory in their balsamic time. Each EO went to the test in vitro (contact toxicity) against V. destructor. Fifty adult female mites, five for each EO and the positive and negative control, were used in each experimental replicate. The positive controls comprised five individuals treated to Amitraz dilute in acetone, and the negative controls included five individuals exposed to acetone alone. To create the working solution to be tested (50 μL/tube), the EOs were diluted (0.5 mg/mL, 1 mg/mL, 2 mg/mL and 4 mg/mL) in HPLC-grade acetone. After 1 h of exposure, mite mortality was manually assessed. Origanum vulgare subsp. viridulum, Thymus capitatus and Thymus longicaulis were the EOs with the highest levels of efficiency at 2 mg/mL, neutralizing (dead + inactivated), 94%, 92% and 94% of parasites, respectively. Salvia rosmarinus EO gave a lower efficacy, resulting in a percentage of 38%. Interestingly, no adverse effects were highlighted in toxicity tests on honeybees. These results show that these OEs of the Lamiaceae family have antiparasitic action on V. destructor. Therefore, they could be used, individually or combined, to exploit the synergistic effect for a more sustainable control of this parasite mite in honeybee farms. Full article

Insect meal as a protein source has been considered a sustainable way to feed animals. H. illucens and T. molitor larvae meal are considered high-protein sources for poultry, also presenting considerable amounts of fatty acids, vitamins, and minerals. However, other potential components in insect meal and insect oil have been more extensively studied in recent years. Chitin, lauric acid, and antimicrobial peptides can present antimicrobial and prebiotic functions, indicating that low levels of their inclusion in insect meal can beneficially affect broilers’ health and immune responses. This systematic review was developed to study the impact of insect products on the health parameters of broilers, and a metanalysis was conducted to evaluate the effects on performance. A database was obtained based on a selection of manuscripts from January 2016 to January 2023, following the mentioned parameters. Both H. illucens and T. molitor meal or oil products had positive effects on poultry health status, especially on the ileal and cecal microbiota population, immune responses, and antimicrobial properties. The average daily gain was greater in broilers fed T. molitor meal compared to H. illucens meal (p = 0.002). The results suggest that low levels of insect meal are suitable for broilers, without resulting in negative effects on body weight gain and the feed conversion ratio, while the insect oil can totally replace soybean oil without negative impacts. Full article

Basophilia is a rare hematologic finding in dogs. This research aimed to describe the hematologic and clinical characteristics of dogs with moderate-to-marked basophilia. CBC reports with blood smear examinations from dogs presented to the North Carolina State University Veterinary Teaching Hospital were retrospectively reviewed for basophilia (>193 cells/µL). We classified basophilia as moderate when counts were ≥500 cells/µL and marked when they reached ≥1000 cells/µL. We compared the hematologic and clinical profiles of dogs with moderate-to-marked basophilia (the basophilia group) to those without basophilia, serving as our control group. In addition, we investigated differences between dogs with marked basophilia versus those with moderate basophilia, as well as between dogs in the basophilia group with and without concurrent eosinophilia. Diseases associated with moderate-to-marked basophilia included eosinophilic lung disease (p < 0.0001), leukemia/myeloproliferative neoplasms (p = 0.004), parasitic infection (p = 0.004), mast cell tumor (p = 0.005), and inflammatory bowel disease (p = 0.02). Overall, dogs with marked basophilia had a lower frequency of inflammatory diseases (51% vs. 70%, p = 0.009) and a higher frequency of neoplastic diseases (48% vs. 26%, p = 0.003) compared to those with moderate basophilia. In the basophilia group, concurrent eosinophilia was only seen in 36% of dogs. Dogs with concurrent eosinophilia were more often diagnosed with inflammatory diseases (77% vs. 58%, p = 0.006), with fewer diagnoses of neoplasia (19% vs. 40%, p = 0.001), compared to dogs without concurrent eosinophilia. The findings of this study offer veterinary clinicians valuable guidance in determining diagnostic priorities for dogs with moderate-to-marked basophilia. Full article

Subclinical mastitis is a common disease that threatens the welfare and health of dairy cows and causes huge economic losses. Somatic cell count (SCC) is the most suitable indirect index used to evaluate the degree of mastitis. To explore the relationship between SCC, diversity in the microbiome, and subclinical mastitis, we performed next-generation sequencing of the 16S rRNA gene of cow’s milk with different SCC ranges. The data obtained showed that the microbiota was rich and coordinated with SCC below 2 × 10⁵. SCC above 2 × 10⁵ showed a decrease in the diversity of microbial genera. When SCC was below 2 × 10⁵, the phylum Actinobacteriota accounted for the most. When SCC was between 2 × 10⁵ and 5 × 10⁵, Firmicutes accounted for the most, and when SCC exceeded 5 × 10⁵, Firmicutes and Proteobacteria accounted for the most. Pathogenic genera such as Streptococcus spp. were absent, while SCC above 2 × 10⁵ showed a decrease in the diversity of microbial genera. SCC was positively correlated with the percentage of Romboutsia, Turicibacter, and Paeniclostridium and negatively correlated with the percentage of Staphylococcus, Psychrobacter, Aerococcus, and Streptococcus. Romboutsia decreased 6.19 times after the SCC exceeded 2 × 10⁵; the SCC increased exponentially from 2 × 10⁵ to 5 × 10⁵ and above 1 × 10⁶ in Psychrobacter. Analysis of the microbiota of the different SCC ranges suggests that the development of mastitis may not only be a primary infection but may also be the result of dysbiosis in the mammary gland. Full article

Phospholipase C Zeta 1 (PLCZ1) is considered a major sperm-borne oocyte activation factor. After gamete fusion, PLCZ1 triggers calcium oscillations in the oocyte, resulting in oocyte activation. In assisted fertilization, oocyte activation failure is a major cause of low fertility. Most cases of oocyte activation failures in humans related to male infertility are associated with gene mutations and/or altered PLCZ1. Consequently, PLCZ1 evaluation could be an effective diagnostic marker and predictor of sperm fertilizing potential for in vivo and in vitro embryo production. The characterization of PLCZ1 has been principally investigated in men and mice, with less known about the PLCZ1 impact on assisted reproduction in other species, such as cattle and horses. In horses, sperm PLCZ1 varies among stallions, and sperm populations with high PLCZ1 are associated with cleavage after intracytoplasmic sperm injection (ICSI). In contrast, bull sperm is less able to initiate calcium oscillations and undergo nuclear remodeling, resulting in poor cleavage after ICSI. Advantageously, injections of PLCZ1 are able to rescue oocyte failure in mouse oocytes after ICSI, promoting full development and birth. However, further research is needed to optimize PLCZ1 diagnostic tests for consistent association with fertility and to determine whether PLCZ1 as an oocyte-activating treatment is a physiological, efficient, and safe method for improving assisted fertilization in cattle and horses. Full article

Leptospirosis is a potentially fatal zoonotic disease caused by infection with pathogenic Leptospira spp. We described reported clinical cases of canine leptospirosis in the council areas of the Inner West and the City of Sydney, Australia, from December 2017 to January 2023 and tested the association with urban spatial (landscape and socioeconomic factors, community seroprevalence, and urban heat island effect) and temporal (precipitation and minimum and maximum temperature) factors and the cases using log-transformed Poisson models, spatially stratified population-adjusted conditional logistic models, General Additive Models (GAMs), and Autoregressive Integrated Moving Average (ARIMA) models. The results suggested that canine leptospirosis is now endemic in the study area. A longer distance to the nearest veterinary hospital (RR 0.118, 95% CI −4.205–−0.065, p < 0.05) and a mildly compromised Index of Economic Resources (IER) (RR 0.202, 95% CI −3.124–−0.079, p < 0.05) were significant protective factors against leptospirosis. In areas proximal to the clinical cases and seropositive samples, the presence of tree cover was a strong risk factor for higher odds of canine leptospirosis (OR 5.80, 95% CI 1.12–30.11, p < 0.05). As the first study exploring risk factors associated with canine leptospirosis in urban Sydney, our findings indicate a potential transmission from urban green spaces and the possibility of higher exposure to Leptospira—or increased case detection and reporting—in areas adjacent to veterinary hospitals. Full article

The aim of this study was to objectively evaluate lameness in dogs affected by a unilateral cranial cruciate ligament rupture (CrCLR) treated with porous tibial tuberosity advancement before surgery and at three different timepoints after surgery, using the GAITRite^® system (version 4.9Wr), a pressure-sensitive walkway system that is able to calculate several spatiotemporal gait parameters simultaneously for each limb. The dogs walked on the pressure-sensitive walkway before (T0) and 30 (T1), 90 (T2), and 120 (T3) days after surgery. Pressure measurements (gait lameness score and total pressure index %) were collected for S (treated with porous TTA) and C (healthy contralateral limb) at T0, T1, T2, and T3 and statistically evaluated. An ANOVA test was performed to compare the data, and a value of p < 0.05 was considered significant. Twenty dogs (n = 20) of various common breeds and ages with CrCLR were enrolled in the study. The results showed that there was a statistically significant difference in the GAIT4Dog^® lameness score (GLS) and TPI% between S and C for each timepoint. Statistically significant differences in the GLS and TPI% between S at T0 and S at T2 and between S at T0 and S at T3 (p < 0.001) were found. The results showed that there was a statistically significant difference in the GAIT4Dog^® lameness score (GLS) and TPI% between S and C for each timepoint. Statistically significant differences in the GLS and TPI% between S at T0 and S at T2 and between S at T0 and S at T3 were found. The GLS and TPI% increased statistically significantly from 90 days after surgery compared to the preoperative measurements. Moreover, comparing the GLS and TPI% between the treated limb and the control limb showed that a statistically significant difference remained at each timepoint. Full article

This study assessed the impact of a mixture of garlic (Allium sativum) and oregano (Origanum vulgare) essential oils (EOGOs) on in vitro dry matter digestibility (IVDMD) and in vivo apparent nutrient digestibility. Different EOGO inclusion levels were evaluated to assess the dose response and potential effects of the mixture. Three EOGO inclusion levels (0.5, 0.75, and 1 mL/kg of incubated dry matter) were evaluated in vitro, while four treatments (0.5, 0.75, and 1 mL/day of EOGO and a control group) were tested in vivo on 12 West African sheep. A randomized controlled trial was conducted using a 4 × 4 design. Blood parameters (glucose, blood urea nitrogen, and β-hydroxybutyrate) were measured to observe the effect of EOGO on the metabolism. The results showed that the inclusion of EOGO significantly enhanced IVDMD at low levels (p < 0.052) compared with the highest levels in treatments containing 0.5 and 0.75 mL/kg of EOGO dry matter. A higher intake of dry matter (DM), crude protein (CP), and neutral detergent fiber (NDF) (p < 0.05) was observed in the in vivo diets with the inclusion of EOGO. In terms of in vivo apparent digestibility, significant differences were found among treatments in the digestibility coefficients of DM, CP, and NDF. EOGO inclusion increased the digestibility of DM. CP digestibility displayed a cubic effect (p < 0.038), with the lowest values of digestibility observed at 1 mL EOGO inclusion. Additionally, NDF digestibility showed a cubic effect (p < 0.012), with the highest value obtained at 0.75 mL of EOGO inclusion. The inclusion levels above 0.75 mL EOGO showed a cubic effect, which indicates that higher concentrations of EOGO may not be beneficial for the digestibility of CP and NDF. Although no significant difference was observed in total digestible nutrients, a linear trend was observed (p < 0.059). EOGO improved the intake of DM, CP, and NDF. EOGO supplementation improved the digestibility of DM and NDF, with optimal levels observed at 0.5 mL/day. No significant effects were observed in the blood parameters. These results suggest that EOGO has the potential as an additive in ruminal nutrition to improve food digestibility and serve as an alternative to antibiotic additives. The use of EOGO potentially improves fiber digestion and may reduce the use of antibiotics in livestock production. Garlic (A. sativum) and oregano (O. vulgare) essential oils effectively modulated fiber digestibility at 0.75 mL/day. Garlic (A. sativum) and oregano (O. vulgare) essential oils have the potential to improve digestibility at low inclusion levels and serve as an alternative to antibiotic additives. The effectiveness of essential oils is greater in a mixture and at lower doses. Full article

Giardia duodenalis infections are common in dogs and are mainly caused by assemblages C and D. The aim of this study was to investigate the correlation between the presence of Giardia duodenalis and different Giardia assemblages detected in symptomatic and asymptomatic dogs and the occurrence of certain clinical signs. All the dogs included (n = 82) were clinically examined, and fecal samples were examined for other parasites and Clostridium spp. Also, G. duodenalis assemblages were detected and the occurrence of certain clinical signs was assessed. A total of 42/82 (51.2%) dogs were symptomatic and had one or more gastrointestinal signs, and 40/82 (48.8%) dogs were asymptomatic. G. duodenalis was found in 25/82 (30.5%) dogs: assemblage C in 10/25 (40%) and assemblage D in 15/25 (60%). Only Cryptosporidium spp. showed a higher coinfection rate with G. duodenalis but that did not have an influence on clinical sign appearance. There was no correlation between the presence of different assemblages of G. duodenalis and the sex of the host or the duration and appearance of certain clinical signs, except the presence of mucus in feces, which was more frequent in dogs invaded with G. duodenalis assemblage C. Further research of other assemblages is needed. Full article

Avipoxvirus (APV), a linear dsDNA virus belonging to the subfamily Chordopoxvirinae of the family Poxviridae, infects more than 278 species of domestic and wild birds. It is responsible for causing avian pox disease, characterized by its cutaneous and diphtheric forms. With a high transmission capacity, it can cause high economic losses and damage to the ecosystem. Several diagnostic methods are available, and bird vaccination can be an effective preventive measure. Ten APV-positive samples were analyzed to update the molecular characterization and phylogenetic analysis of viruses isolated in Portugal between 2017 and 2023. A P4b gene fragment was amplified using a PCR, and the nucleotide sequence of the amplicons was determined using Sanger sequencing. The sequences obtained were aligned using ClustalW, and a maximum likelihood phylogenetic tree was constructed. With this study, it was possible to verify that the analyzed sequences are distributed in subclades A1, A2, B1, and B3. Since some of them are quite similar to others from different countries and obtained in different years, it is possible to conclude that there have been several viral introductions in Portugal. Finally, it was possible to successfully update the data on Avipoxviruses in Portugal. Full article

Bidirectional patellar luxation (BPL) is a relatively rare form of patellar luxation, with limited information reported regarding breed predisposition and etiology. The purpose of this study was to describe the patient characteristics and radiographic measures of proximodistal patellar position associated with BPL in dogs in Japan, compared with dogs with medial patellar luxation (MPL). A retrospective medical record search of surgically corrected MPL and BPL dogs was performed, and breed, age, sex, body weight, and presence of the patella alta in the extended-stifle position were recorded. The ratio of the patellar ligament length to patella length (PLL/PL) and the ratio of the distance between the proximal pole of the patella and the femoral condyle to patella length (A/PL) were measured on stifle radiographs. A total of 35 dogs with BPL and 95 dogs with MPL were included. The BPL was most commonly present in Toy Poodles (odds ratio compared to MPL dogs: 7.05) in the present study. There were no significant differences in age, sex, or body weight between the BPL and MPL groups. Patella alta in the extended-stifle position was more common in the BPL group (23.4%) than in the MPL group (0.8%). However, there were no significant differences in PLL/PL or A/PL between the BPL and MPL groups. The study highlights BPL in different dog breeds in Japan, and suggests that the occurrence of BPL may be related to stifle extension. However, more research is needed to fully understand the etiology of BPL. Full article

In horses, the prevalence of obesity is high and associated with serious metabolic pathologies. Being a broodmare has been identified as a risk factor for obesity. In other species, maternal obesity is known to affect the development of the offspring. This article is a follow-up study of previous work showing that Obese mares (O, n = 10, body condition score > 4.25 at insemination) were more insulin resistant and presented increased systemic inflammation during pregnancy compared to Normal mares (N, n = 14, body condition score < 4 at insemination). Foals born to O mares were more insulin-resistant, presented increased systemic inflammation, and were more affected by osteoarticular lesions. The objective of the present study was to investigate the effect of maternal obesity on placental structure and function, as well as the fatty acid profile in the plasma of mares and foals, colostrum, and milk until 90 days of lactation, which, to our knowledge, has been poorly studied in the horse. Mares from both groups were fed the same diet during pregnancy and lactation. During lactation, mares were housed in pasture. A strong heat wave, followed by a drought, occurred during their 2nd and 3rd months of lactation (summer of 2016 in the Limousin region, France). In the present article, term placental morphometry, structure (stereology), and gene expression (RT-qPCR, genes involved in nutrient transport, growth, and development, as well as vascularization) were studied. Plasma of mares and their foals, as well as colostrum and milk, were sampled at birth, 30 days, and 90 days of lactation. The fatty acid composition of these samples was measured using gas chromatography. No differences between the N and O groups were observed for term placental morphometry, structure, or gene expression. No difference in plasma fatty acid composition was observed between groups in mares. The plasma fatty acid profile of O foals was more pro-inflammatory and indicated an altered placental lipid metabolism between birth and 90 days of age. These results are in line with the increased systemic inflammation and altered glucose metabolism observed until 18 months of age in this group. The colostrum fatty acid profile of O mares was more pro-inflammatory and indicated an increased transfer and/or desaturation of long-chain fatty acids. Moreover, O foals received a colostrum poorer in medium-chain saturated fatty acid, a source of immediate energy for the newborn that can also play a role in immunity and gut microbiota development. Differences in milk fatty acid composition indicated a decreased ability to adapt to heat stress in O mares, which could have further affected the metabolic development of their foals. In conclusion, maternal obesity affected the fatty acid composition of milk, thus also influencing the foal’s plasma fatty acid composition and likely participating in the developmental programming observed in growing foals. Full article

Direct-targeted next-generation sequencing (tNGS), with its undoubtedly superior diagnostic capacity over real-time PCR (RT-PCR), and direct-non-targeted NGS (ntNGS), with its higher capacity to identify and characterize multiple agents, are both likely to become diagnostic methods of choice in the future. tNGS is a rapid and sensitive method for precise characterization of suspected agents. ntNGS, also known as agnostic diagnosis, does not require a hypothesis and has been used to identify unsuspected infections in clinical samples. Implemented in the form of multiplexed total DNA metagenomics or as total RNA sequencing, the approach produces comprehensive and actionable reports that allow semi-quantitative identification of most of the agents present in respiratory, cloacal, and tissue samples. The diagnostic benefits of the use of direct tNGS and ntNGS are high specificity, compatibility with different types of clinical samples (fresh, frozen, FTA cards, and paraffin-embedded), production of nearly complete infection profiles (viruses, bacteria, fungus, and parasites), production of “semi-quantitative” information, direct agent genotyping, and infectious agent mutational information. The achievements of NGS in terms of diagnosing poultry problems are described here, along with future applications. Multiplexing, development of standard operating procedures, robotics, sequencing kits, automated bioinformatics, cloud computing, and artificial intelligence (AI) are disciplines converging toward the use of this technology for active surveillance in poultry farms. Other advances in human and veterinary NGS sequencing are likely to be adaptable to avian species in the future. Full article

Porcine circovirus type 3 (PCV3) is commonly associated with clinical symptoms such as porcine dermatitis and nephropathy syndrome (PDNS)-like lesions, respiratory signs, and reproductive disorders. This study aimed to investigate the epidemiology of PCV3 in a boar stud. The objectives were to detect PCV3 in semen, as well as matched serum, oral fluid, and preputial fluid samples from adult boars using quantitative polymerase chain reaction (qPCR), analyze PCV3-IgG antibody data, and genetically characterize a positive sample. A total of 112 samples from 28 boars were collected from a large-scale pig farm in Guangxi, China. The qPCR results showed that the PCV3 DNA was not detected in semen, with a positive rate of 0% (0/28), while it was detected in serum (3.57%—1/28), oral fluid (64.28%—18/28), and preputial fluid (46.4%—13/28). The seropositivity rate of PCV3-IgG in serum was 82.14% (23/28) according to the indirect enzyme-linked immunosorbent serologic assay (ELISA) results. Phylogenetic analysis revealed that one of the PCV3 isolates belonged to the PCV3c clades. This is the first report of PCV3 detection in preputial fluid from boars. The results suggest that PCV3 is transmitted among boars on pig farms and exhibits epidemic characteristics. Full article

This paper describes cases of urolithiasis in fattening pigs on two farms (A and B). Bladder rupture due to urethral obstruction with calculi was the principal finding during the necropsy of the pigs. An in-depth diagnostic examination was performed to elucidate possible pathophysiological mechanisms, namely Fourier-transform infrared spectrophotometry (FT-IR) analysis of the uroliths, blood analysis (farm A: 5 samples, farm B: 10 samples) for assessing concentrations of minerals, the bone resorption marker cross-linked C-telopeptide of type 1 collagen (CTX), parathyroid hormone (PTH), and vitamin D components, biochemical urinalysis (farm A: 5 samples, farm B: 7 samples), microscopic examination of urinary sediment (Farms A and B: 7 samples each), mineral composition of the feed, and analysis of the drinking water. Calcium carbonate was the main component found in stones from both farms, and calcium carbonate and struvite were the main components found in crystals from farms A and B, respectively. On farm A, urinary calcium excretion and urinary pH were high; on farm B, urinary phosphorus was high and urinary calcium was low with a normal urinary pH. The mineral compositions of the feed and drinking water were similar on both farms and could therefore not explain the difference between the two farms. Disturbances in calcium and phosphorus absorption and homeostasis might have been involved in these problems. Further research should focus on the calcium, phosphorus, and vitamin D levels in the feed and take into account other factors, such as the absorption and excretion of minerals due to gut and urinary microbiota. Full article

Background: ACVIM developed and published guidelines for staging myxomatous mitral valve degeneration in dogs in 2009. An updated version was published in 2019. The present study aimed to investigate whether these guidelines are actually used by the intended public more than a decade after their first publication. Methods: An online survey was distributed among Dutch and Belgian veterinarians through online channels and mailing lists. Results: Of the 524 responses, only 363 complete surveys were analyzed. The ACVIM guidelines are used by 60% of the respondents. Veterinarians find it more difficult to differentiate stage B1 from B2 in asymptomatic dogs compared to diagnosing stage C. Three-quarters of the respondents would recommend echocardiography for an incidentally detected new murmur with an intensity of 3 out of 6 in an adult dog. Two-thirds of the respondents find coughing a convincing finding for stage C disease. Close to half of the respondents associate a horizontal, dull percussion line with pulmonary edema. For confirming cardiogenic pulmonary edema, 98% of the respondents used thoracic radiographs. Conclusions: Veterinary practitioners might not have the expected training and equipment to be able to apply the guidelines in their practices, especially in the differentiation of stage B1 from stage B2. Full article

Platelet concentrates (PCs) have become widely used in veterinary and human medicine. The PCs consist mainly of supraphysiological concentrations of platelets and, therefore, growth factors (GFs) which are stored within platelet α-granules. Among PCs, Platelet-Rich Plasma (PRP) is characterised by low-density fibrin. Research on the effect of PCs in cattle has surged in recent years; in particular, evidence has shown the positive use of PRP for treating reproductive problems, in vitro production of bovine embryos, sole ulcers and udder diseases. The aim of this report is to critically review, in accordance with the PRISMA guidelines, the available literature reporting clinical application in the bovine practice of PRP. Three bibliographic databases PubMed, Web of Science and Scopus were used for a broad search of “platelet concentrates” OR “PRP” OR “platelet-rich plasma” OR “PRF” OR “platelet-rich fibrin” AND “cows” OR “cattle”. From 1196 papers, only six met the inclusion criteria. Two papers described the use of PRP in mastitis, two papers in uterine dysfunction and two papers in ovarian dysfunction. PRP offered a low-cost, easily obtained therapeutic option and showed positive results for these patients. However, given the different pathologies and definitions involved, further studies are necessary to assess its full clinical potential. Full article

The most important pathogenic Mycoplasma species in bovines are Mycoplasma bovis (M. bovis) and Mycoplasma mycoides subsp. mycoides (Mmm). Mmm causes contagious bovine pleuropneumonia (CBPP), which is a severe respiratory disease widespread in sub-Saharan Africa but eradicated in several countries, including China. M. bovis is an important cause of the bovine respiratory disease complex (BRD), characterized worldwide by pneumonia, arthritis, and mastitis. Secreted proteins of bacteria are generally considered virulence factors because they can act as toxins, adhesins, and virulent enzymes in infection. Therefore, this study performed a comparative proteomic analysis of the secreted proteins of M. bovis and Mmm in order to find some virulence-related factors as well as discover differential diagnostic biomarkers for these bovine mycoplasmas. The secretome was extracted from both species, and liquid chromatography-tandem mass spectrometry was used, which revealed 55 unique secreted proteins of M. bovis, 44 unique secreted proteins of Mmm, and 4 homologous proteins. In the M. bovis secretome, 19 proteins were predicted to be virulence factors, while 4 putative virulence factors were identified in the Mmm secretome. In addition, five unique secreted proteins of Mmm were expressed and purified, and their antigenicity was confirmed by Western blotting assay and indirect ELISA. Among them, Ts1133 and Ts0085 were verified as potential candidates for distinguishing Mmm infection from M. bovis infection. Full article

Ticks pose a major threat to cattle health and production in Pakistan because they transmit pathogens of diseases like Babesiosis and Theileriosis. Hyalomma spp., found across Africa, Asia, and Europe, are especially problematic. This study explored biocontrol of Hyalomma spp. using spore-free fungal culture filtrates collected from dairy farm soil in Kohat, Pakistan. Three fungal species of the genera Alternaria, Aspergillus, and Penicillium were isolated, and their filtrates were tested against tick adults and larvae. Filtrate concentrations were prepared at different strengths. Data were taken after the exposure of adults and larvae ticks to various concentrations of the fungal filtrates. Results indicated that at 100% concentration, all fungal filtrates induced 100% mortality in adults and larvae. Decreasing filtrate concentration lowered tick mortality. The lowest concentration caused the least mortality. The effect was time- and dose-dependent. In conclusion, spore-free fungal culture filtrates can provide biocontrol of Hyalomma spp. in a time- and concentration-dependent manner. Further research should explore the active compounds causing mortality and optimal application methods. The process outlined here provides a natural biocontrol alternative to chemical pesticides to reduce tick infestations and associated cattle diseases in Pakistan. Full article

Ultrasonography is a valuable diagnostic tool extensively used in the andrology of human and domestic animals, including dogs. This review aims to provide an overview of various technologies based on ultrasound, from the basic B-Mode ultrasonography to the more recent advancements, such as contrast-enhanced ultrasonography (CEUS) and ultrasound elastography (UEl), all of which are utilized in the evaluation of canine testicles. The review outlines the principles behind each of these technologies and discusses their application in assessing normal and abnormal testicular conditions. B-mode canine testicular ultrasonography primarily focuses on detecting focal lesions but has limitations in terms of objectivity. Other technologies, including Doppler ultrasonography, B-Flow, and CEUS, allow for the characterization of vascular patterns, which could be further measured using specific applications like spectral Doppler or quantitative CEUS. Additionally, ultrasound elastography enables the assessment of parenchyma stiffness both qualitatively and quantitatively. These ultrasound-based technologies play a crucial role in andrology by providing valuable information for evaluating testicular function and integrity, aiding in the identification of pathological conditions that may impact the health and quality of life of male dogs. Full article

Surgical techniques are gaining attention for treating physical diseases in aquaculture and aquarium fish. Sturgeon is a suitable species for surgical experiments due to its industrial significance. Maintaining homeostasis is crucial during surgical procedures, and the liver plays a major role in immune regulation. High temperature is suggested to improve physiological activity and wound healing. This study investigated differences in hepatectomy sturgeons’ tolerance and histopathological responses of internal organs. Moreover, this study investigated the effects of high temperatures on wound healing and hematopoietic recovery in fish undergoing surgical procedures. The liver condition was found to play a pivotal role in the analysis, and cortisol levels were affected by anesthesia. The results showed that high temperature facilitated hematopoietic recovery and wound healing, but excessive induction of physiological activity caused damage. Managing high temperatures and liver conditions induced a remarkable improvement in wound healing. However, anesthesia itself can be a significant stressor for fish, and wound healing requires a greater amount of energy. Further research is needed to understand the stress factors caused by surgical procedures and anesthesia and to promote animal welfare in fishery products. Full article

Prototheca infections are rare in cats, and they are usually associated with cutaneous or subcutaneous infections by P. wickerhamii, with no evidence of neurological signs or systemic disease. In this study, we report the identification of prototheca in the cerebrospinal fluid (CSF) of a cat with neurological symptoms. Fourteen CSF samples were gathered from cats presented with neurological disease between 2012 and 2014. The inclusion criteria for the samples were an increase in CSF protein and cell number (pleocytosis), suggestive of an infectious inflammatory status of the central nervous system (CNS). Nine samples fulfilled the inclusion criteria (inflammatory samples), while five samples, used as control, did not (non-inflammatory samples). All the samples were screened molecularly for different pathogens associated with CNS disease in cats, including prototheca. Out of 14 CSF samples, only one inflammatory sample tested positive for prototheca. Upon sequence and phylogenetic analysis of the amplicon, the strain was characterized as P. bovis. This report is the first documented evidence of prototheca in the cerebrospinal fluid of a cat with neurological signs. Prototheca should be considered in the diagnostics procedures on the CNS of cats presented with infectious diseases. Full article

Antimicrobial resistance is a widespread global health problem. The presence of resistant bacteria and antibiotic resistance genes has been demonstrated not only in humans but also in animals, including pets. Stray cats share the urban environment with people and pets. This may facilitate transmission of resistant bacteria and resistance genes between stray animals, people and domestic animals. Several studies have investigated the role of stray cats as a fecal carrier of ESBL-producing bacteria. However, there are many genes and resistance mechanisms that can be detected in commensal E. coli, which, because of its genetic plasticity, is considered an indicator for monitoring antibiotic resistance. In this study, rectal swabs were collected from stray cats from colonies and shelters in the city of Monza (Monza Brianza, Italy) to isolate commensal E. coli. Phenotypic tests, such as the minimum inhibitory concentration (MIC) and the double disc test (DDST), and molecular analyses to detect antimicrobial resistance genes (ARGs) were used to study the resistance of these isolates. The results obtained confirm that stray cats can carry ESBL-producing E. coli (6.7%) and genes conferring resistance to other important antibiotic classes such as tetracyclines and sulfonamides. Full article

Rumen microbial protein synthesis (MPS) provides at least half of the amino acids for the synthesis of milk and meat protein in ruminants. As such, it is fundamental to global food protein security. Estimating microbial protein is central to diet formulation, maximising nitrogen (N)-use efficiency and reducing N losses to the environment. Whilst factors influencing MPS are well established in vitro, techniques for in vivo estimates, including older techniques with cannulated animals and the more recent technique based on urinary purine derivative (UPD) excretion, are subject to large experimental errors. Consequently, models of MPS used in protein rationing are imprecise, resulting in wasted feed protein and unnecessary N losses to the environment. Newer ‘omics’ techniques are used to characterise microbial communities, their genes and resultant proteins and metabolites. An analysis of microbial communities and genes has recently been used successfully to model complex rumen-related traits, including feed conversion efficiency and methane emissions. Since microbial proteins are more directly related to microbial genes, we expect a strong relationship between rumen metataxonomics/metagenomics and MPS. The main aims of this review are to gauge the understanding of factors affecting MPS, including the use of the UPD technique, and explore whether omics-focused studies could improve the predictability of MPS, with a focus on beef cattle. Full article

This study aims to evaluate the effectiveness of the dietary addition of Spirulina platensis (SP) and/or garlic powder (GP) on heat-stressed broiler chickens. For this purpose, 600 Ross-308 broiler chicks were allocated at 22 days of age into five groups (G1–G5), each comprising six groups of 20 birds each. Chickens kept in G1 (negative control) were fed a basal diet and raised at 26 ± 1 °C. Chickens kept in G2 to G5 were exposed to periodic heat stress (35 ± 1 °C for 9 h/day) from 22 to 35 days old. Chickens in G2 (positive control) were provided a basal diet, while G3, G4, and G5 were fed a basal diet enriched with SP (1 g/kg diet), GP (200 mg/kg diet), or SP/GP (1 g SP/kg + 200 mg GP/kg diet), respectively. The assessment parameters included the chickens’ performance, malondialdehyde and total antioxidant capacity, blood biochemistry, intestinal morphology, and modulation of lactobacilli and total coliforms in the intestinal microbiota. Our findings demonstrated that supplementing heat-stressed chickens with SP and/or GP significantly mitigated the negative effects on the European production efficiency index (EPEF), survival rate, cholesterol profile, and oxidative stress markers. Chickens supplemented with GP and/or SP exhibited significantly better EPEF and survivability rates. Heat stress had a significant impact on both the gut structure and gut microbiota. However, SP and/or GP supplementation improved the gut morphology, significantly increased the intestinal lactobacilli, and reduced the coliform contents. It was also found that the simultaneous feeding of SP and GP led to even higher recovery levels with improved lipid metabolites, immunity, and oxidative status. Overall, supplementing chickens with SP and/or GP can alleviate the negative effects of heat stress. Full article