Vet. Sci., Volume 9, Issue 2 (February 2022) – 63 articles

The emergence of resistance to chemical drugs in beekeeping is becoming a phenomenon of widespread concern. One promising alternative to the use of chemicals is entomopathogenic organisms that are environmentally friendly and are capable of stopping the expression of resistance once it has evolved. In the recent past, the scientific community has carried out several experiments addressing the use of microbiological control agents. In particular, experimental studies using entomopathogenic fungi have had more success in honey bee research. With their adherence properties and their ability to digest the cuticle and overcome the host defense mechanism, they could be a suitable ingredient in bioacaricides. Several promising fungi have been identified in the search for effective means to control pest populations. The data obtained from the different experiments are interesting and often favorable to their use, but there are also conflicting results. The aim of this review is to describe the state of the art on the topic under investigation. Full article

Equine melanocytic neoplasm (EMN) is a common disease in older grey horses. The purpose of this study was to examine the potential proteins throughout EMN stages from faecal proteomic outlining using functional analysis. Faecal samples were collected from the rectum of 25 grey horses divided into three groups; normal group without EMN (n = 10), mild EMN (n = 6) and severe EMN (n = 9). Based on the results, 5910 annotated proteins out of 8509 total proteins were assessed from proteomic profiling. We observed differentially expressed proteins (DEPs) between the normal group and the EMN group, and 109 significant proteins were obtained, of which 28 and 81 were involved in metabolic and non-metabolic functions, respectively. We found 10 proteins that play a key role in lipid metabolism, affecting the tumour microenvironment and, consequently, melanoma progression. Interestingly, FOSL1 (FOS like 1, AP-1 transcription factor subunit) was considered as a potential highly expressed protein in a mild EMN group involved in melanocytes cell and related melanoma. Diacylglycerol kinase (DGKB), TGc domain-containing protein (Tgm2), structural maintenance of chromosomes 4 (SMC4) and mastermind-like transcriptional coactivator 2 (MAML2) were related to lipid metabolism, facilitating melanoma development in the severe-EMN group. In conclusion, these potential proteins can be used as candidate biomarkers for the monitoring of early EMN, the development of EMN, further prevention and treatment. Full article

Herbal products such as essential oils may play a promising role in the treatment of infections caused by gastrointestinal nematodes (GINs). The aim of this study was to evaluate the in vitro potential of 11 essential oils (EOs) and one binary combination of isolated EO compounds, as well as the in vivo anthelmintic efficacy of two EO formulations. Four GIN genera were identified in the coproculture examination: Haemonchus, Trichostrongylus, Teladorsagia and Chabertia. The in vitro egg hatch test (EHT) was performed at six different concentrations (50, 12.5, 3.125, 0.781, 0.195 and 0.049 mg/mL) for each EO, whereas in the in vivo faecal egg count reduction test (FECRT), each EO sample was diluted in sunflower oil and orally administrated at a dose of 100 mg/kg to the different group of animals. In the EHT, the EOs of Origanum vulgare, Foeniculum vulgare, Satureja montana, Satureja hortensis and two types of Thymus vulgaris were the most effective. The dominant compounds of these EOs were carvacrol, thymol, anethol, p-cymene and γ-terpinene, indicating their importance for the anthelmintic activity. In the FECRT, both T. vulgaris EO type 1 and linalool:estragole combination show an anthelmintic potential with a mean effect on FECR of approximately 25%. The results suggest the possible role of tested EOs as anthelmintic agents in sheep farms, although further in vivo tests are needed. Full article

Zebrafish are widely used to investigate candidate genes for human diseases. While the emergence of CRISPR-Cas9 technology has revolutionized gene editing, the use of individual guide RNAs limits the efficiency and application of this technology in functional genetics research. Multiplexed genome editing significantly enhances the efficiency and scope of gene editing. Herein, we describe an efficient multiplexed genome editing strategy to generate zebrafish mutants. Following behavioural tests and histological examination, we identified one new candidate gene (tmem183a) for hearing loss. This study provides a robust genetic platform to quickly obtain zebrafish mutants and to identify candidate genes by phenotypic readouts. Full article

Cryptosporidium cuniculus is a zoonotic parasite responsible for cryptosporidiosis cases and outbreaks in both humans and rabbits. Since there are no molecular Cryptosporidium spp. infection data in rabbits (Oryctolagus cuniculus) from Spain, our aim was to gather information about this parasite in wild European rabbits from Tenerife, Canary Islands (Spain). A total of 100 faecal samples were collected from rabbits from eight municipalities of Tenerife. Microscopic analysis showed that 4.0% of the samples presented structures compatible with Cryptosporidium oocyst. A nested polymerase chain reaction (PCR) targeting 18S ribosomal RNA (rRNA) gene fragments was carried out, and sequencing confirmed the identity of C. cuniculus in one sample (1.0%). The sample was successfully subtyped using nested PCR analysis of the 60-kDa glycoprotein (gp60) gene as the subtype VbA26R3. This study confirms the presence of C. cuniculus in wild rabbits from Tenerife, providing new information on the occurrence of this zoonotic parasite. Further studies are required to better understand the epidemiology of Cryptosporidium spp. in wild rabbits in Spain and their possible public health repercussions. Full article

The study investigated whether the repeat use of needles used to inject piglets with iron, influences the force required to puncture into piglet cadaver skin. Pig units (n = 31) were surveyed on needle reuse and injection practices, and these findings informed subsequent laboratory-based experiments on needle puncture force into piglet cadaver tissues. A 21 G 5/8” needle was reported as the most used needle type (67.7%), with 80.6% reporting needle reuse; 38.7% changed the needle between litters or earlier if damaged, 16.1% every three litters and 22.5% when it felt blunt or damaged, after each injection session or when changing the bottle of iron solution. There was a significant difference in puncture force between the 1st and 36th use (p < 0.05), and between the 1st and 100th use (p = 0.0015), but not between the 1st and 12th or 36th use (p > 0.999 and p = 0.8313, respectively). Scanning electron microscopy (SEM) imaging showed appreciable damage to the needle tip after 12 uses. The repeat use of needles in piglet cadavers increased the force of needle puncture compared to first-time use. When extrapolated to live animals, the use of blunt needles has the potential to cause pain and distress. Full article

Trichotillomania (TTM) is a body-focused repetitive disorder affecting as much as 0.5 to 2% of the population, with women four times more likely to be affected than men. This disorder causes impairment in daily function and significant distress. A potential animal model for this disorder is the inbred C57BL/6J mouse which displays clinical signs and behavioral characteristics similar to those described for people affected by this disorder. Because alcohol-preferring P rats also display similar clinical signs and behavioral characteristics, it was hypothesized that this selectively bred stock could be an additional animal model. In this study, 112 female P rats were recorded on digital media for 15 min after being sprayed with a mist of water and assessed for grooming patterns—oral, manual, and scratching. Significant elevations in scratching and oral grooming behavior were predictive of the future development of skin lesions. These findings suggest that P rats may be an additional model to study TTM, with the advantage of increased genetic variation (i.e., non-inbred) which mirrors the human population. The use of this model may help to identify preventative and therapeutic interventions for humans and other animals with similar body-focused repetitive disorders. Full article

Among the respiratory disorders in dogs from small breeds, tracheal collapse is one of the most commonly found in clinical practice. Presently, acupuncture is widely used as an alternative treatment which was shown to bring about positive effects in the treatment of human respiratory diseases. The present study demonstrated the effect of acupuncture on tracheal collapse dogs. We hypothesized that acupuncture can help dogs suffering from tracheal collapse by improving various parameters including heart rate variability, serum biomarkers for oxidative stress, exercise performance, and quality of life. Twenty client-owned dogs from small breeds with tracheal collapse disease were enrolled. The study was divided into two 5-week periods. During the first period, the dogs received normal veterinary care but received no acupuncture treatment (NAC). After completing that period, all forms of treatment were withheld for one week before the beginning of the second period. In the second period, all dogs restarted normal veterinary care and underwent acupuncture treatment (AC) once a week for five consecutive weeks. Blood was collected at the beginning and end of each of the two periods for malondialdehyde (MDA) level measurement. Heart rate variability (HRV) was recorded at the 1st, 3rd and 5th weeks of both periods. Exercise tests were performed at the beginning and end of AC period and questionnaire interviews with the owners were accomplished at the end of each period. The results showed that acupuncture can alleviate clinical signs of tracheal collapse, reduce MDA level, and improve sympathovagal balance. We suggest that acupuncture treatment could be used as an adjunct treatment for canine tracheal collapse. Full article

Neosporosis is recognized as one of the major causes of bovine abortion worldwide. Canids are the main definitive host for this parasite and the presence of dogs in the farm is an important factor for the Neospora caninum infection in bovines. Since, in the province of Lecce, located in the Apulia region of Southern Italy, there are no studies showing the presence of the infection in farm animals, the objective was to perform a serological evaluation for anti-N. caninum antibodiesin serum from 706 dairy cattle and 21 farm dogs located in 40 farms uniformlydistributed over the territory.The presence of N. caninum infection was confirmed in 90.0% (36/40) of the 40 farms examined. The results obtained on all serum samples by an enzyme-linked immunosorbent assay (ID Screen^®Neospora caninum competition ELISA kit) for anti-N. caninum antibodies showed a seropositivity rate of 21.1% (149/706) among dairy cows, with a statistically significant higher percentage of positive subjects in the animals over two years old and a positivity rate of 42.9% (9/21) in tested dogs. The obtained data confirmed the presence of neosporosis even in the Lecce area, where it could therefore represent an important cause of abortion and economic losses. Full article

Varicocele is a common pathological condition of testis that is related to male fertility problems. A 3-year age Chios ram had an abnormally enlarged scrotal area, was excluded from reproductive duties, and was euthanized with the owners’ permission. The main pathological finding was the presence of bilateral multinodular spermatic cord enlargement with laminated vascular thrombi. Histopathological examination revealed commonly mineralized thrombi within the lumen of veins of the pampiniform plexus, inflammation and testicular degeneration. The epididymides were transported to the laboratory and each cauda region was sliced and washed (8 mL water for injection/epididymis), and the epididymal sperm samples were collected. Sperm motility variables (CASA), viability (eosin-nigrosine), morphology (SpermBlue^®), and DNA integrity (Acridine Orange Test, AOT) were assessed. The total and progressive motility were low in semen samples of both sides (30.00% and 1.00% vs. 42.60% and 2.50% for left and right epididymis, respectively). Low viability values were observed for both sides (26.00% vs. 23.00% for left and right epididymis, respectively), while sperm morphological abnormalities were within normal limits. No sperm with DNA damage were detected. The results of this case report indicate that varicocele is associated with testis dysfunction and degradation of ram semen quality, mainly affecting motility and kinematics. Full article

This sequential explanatory mixed-method study consisted of analytical, cross-sectional, and qualitative studies. The research was conducted in the Khao Nor and Khao Kaew areas of the Banphot Pisai districts of Nakhon Sawan Province in 2019. Here, we examined the rodent contact characteristics of villagers in these areas and determined the potential characteristics/risk factors associated with rodents using a semi-structured questionnaire, key informant interview (KII), and focus group discussion (FGD). Results of the quantitative study (N1 = 372) characterized participants that contacted rodents per gender, age, occupation, knowledge, attitude, and practice (KAP), including their cultural contexts, and beliefs. Ninety participants (24.2%) reported contact with rodents, and the reasons for their direct physical rodent contact were hunting (35, 9.4%), killing (41, 11.0%), preparing rodents as food (33, 8.9%), consuming cooked meats (12, 3.2%), feeding food (4, 1.1%), cleaning feces (17, 4.6%), and cleaning carcasses (33, 8.9%). Moreover, logistic regression results showed that males encountering rodents were statistically significant (Adjusted OR = 3.137, 95% CI 1.914–5.139, p < 0.001). Low monthly household income (<THB 15,000 or <USD 450) was also negatively statistically significant with encountering rodents (Adjusted OR = 0.57, 95% CI 0.33–0.99, p = 0.04). Additionally, the villagers had a low level of knowledge toward zoonotic diseases and inappropriate attitudes and practices toward contacting rodents and zoonotic diseases. Thirty-five qualitative study participants (N2) participated in the KIIs and FGDs. Various rodent contact activities were also reported among the qualitative research participants, such as hunting, consumption, and selling them to their friends and neighbors. However, these rodents also destroyed their belongings, crops, and plants. Some participants also reported that rodents accounted for leptospirosis transmission. As a result, communication intervention should be planned to provide appropriate knowledge and attitude to the villagers, especially among those who have close contact with rodents in the understudied area. Full article

The purpose of the study was to quantify serum and fecal amino acids (AA) in cats with chronic kidney disease (CKD) and compare to healthy cats. Thirty-five cats with International Renal Interest Society Stage 1–4 CKD and 16 healthy mature adult and senior client-owned cats were included in this prospective cross-sectional study. Sera were analyzed for 25 AA concentrations using an ion exchange chromatography AA analyzer with post column ninhydrin derivatization. Voided fecal samples were analyzed for 22 AA concentrations using liquid chromatography with tandem mass spectrometry. CKD cats had lower serum concentrations of phenylalanine (mean difference ± standard error of the mean: 12.7 ± 4.3 µM; p = 0.03), threonine (29.6 ± 9.2 µM; p = 0.03), tryptophan (18.4 ± 5.4 µM; p = 0.005), serine (29.8 ± 12.6 µM; p = 0.03), and tyrosine (11.6 ± 3.8 µM; p = 0.01) and higher serum concentrations of aspartic acid (4.7 ± 2.0 µM; p = 0.01), β-alanine (3.4 ± 1.2 µM; p = 0.01), citrulline (5.7 ± 1.6 µM; p = 0.01), and taurine (109.9 ± 29.6 µM; p = 0.01) when compared to healthy cats. Fecal AA concentrations did not differ between healthy cats and CKD cats. 3-Methylhistidine-to-creatinine did not differ between healthy cats with and without muscle loss. Cats with CKD IRIS Stages 1–4 have a deranged serum amino acid profile compared to healthy cats. Full article

Bovine leptospirosis is an infectious zoonotic disease causing reproductive problems and economic losses in livestock. This work reports, for the first time in Sicily (South Italy), an outbreak of Leptospira interrogans serogroup Pomona that occurred in cattle farms within the Nebrodi Park and was mainly characterized by full-term abortion. Blood and urine samples were collected at different time points from animals of six different farms (Farms A–F) sharing the same grazing area. Research of antibodies against pathogenic Leptospira species in serum samples was carried out via Micro Agglutination Test (MAT). Urine samples were subjected to pathogen isolation and molecular analyses via TaqMan Real Time-PCR. Genotyping of Leptospira species was obtained by Multi-locus sequence typing. MAT detected antibodies against Leptospira interrogans serogroup Pomona in serum samples of all the farms. Pathogenic Leptospira spp. DNA and culture isolation was obtained from urine samples. Genotyping confirmed the excretion of L. interrogans serogroup Pomona. This study describes clinical manifestations, diagnostic implications and epidemiological characteristics of an outbreak in cattle due to L. interrogans Pomona in a protected multi-host area, where domestic and wild animals share the same habitat, suggesting a role of wild species in transmission and persistence of Pomona serogroup among cattle. Full article

Canine mammary tumor (CMT) is the most common tumor in canines after skin tumors. Long noncoding RNAs (lncRNAs) have crucial roles in human breast tumor initiation and progression, but the role of lncRNAs in canine mammary tumors is unclear. We analyzed the expression profiles of canine mammary tumors and their adjacent non-neoplastic tissue to explore abnormally expressed lncRNAs. LncRNA expression was detected by qRT–PCR. After overexpression of lnc40589 and knockdown of lnc34977 in CMT cells, CCK-8, colony formation, wound healing and Transwell assays were used to assess the proliferation, migration and invasive ability of canine mammary tumor cells. We also established a mammary tumor-bearing nude mouse model. GO analysis and KEGG pathway analysis demonstrated that the differentially expressed lncRNAs were closely related to the mammary tumor. lnc40589 was significantly upregulated and lnc34977 was significantly downregulated in CMTs. In addition, lnc40589 inhibits cell proliferation, migration and invasion, while lnc34977 promotes cell proliferation, migration and invasion. In addition, lnc34977 promotes the development of mammary tumors in animals. Taken together, our study results reveal the lncRNA expression profiles in CMTs and indicate that lnc34977 promotes the development of CMT both in cell culture and in vivo. Full article

Feline herpesvirus 1 (FHV-1) is one of the main causes of upper respiratory tract infection in cats. Despite its veterinary importance, no previous studies investigated the occurrence of this virus in Egypt. In the present work, a total number of one hundred forty (N = 140) conjunctival and/or oropharyngeal swabs were collected from symptomatic cats during veterinary clinic visits located in two Egyptian provinces. Virus isolation was performed in the Chorioallantoic membranes (CAMs) of 12-days-old SPF eggs. Interestingly, the embryos showed stunting growth and abnormal feathering and infected CAMs showed edematous thickening and cloudiness with characteristic white opaque pock lesions. Polymerase chain reaction (PCR) amplification of the thymidine kinase gene (TK) was successful in 16/140 (11.4%) of the suspected cases. Two of the amplified genes were sequenced and the TK gene sequences of the FHV-1 isolates were highly similar to other reference strains in the GenBank database. Given the above information, the present study represents the first report of feline herpesvirus type 1 (FHV-1) in domestic cats in Egypt. Further studies on the causes of upper respiratory tract infections in cats as well as vaccine efficacy are needed. Full article

Infection of pig farms with porcine reproductive and respiratory syndrome virus (PRRSV) and porcine circovirus type 2 (PCV2) causes substantial economic losses globally. However, little epidemiological data of PRRSV and PCV2 in the Hong Kong Special Administrative Region (HKSAR) were available. This pilot study aimed to provide baseline information of the prevalences of PPRSV and PCV2 in the HKSAR. A complex survey was conducted from 3 February 2020 to 11 March 2021 on 29 of the 40 pig farms in the HKSAR, with five pigs each from seven age groups (representing key production stages) tested using a real-time PCR. Evidence of presence of PRRSV European strain (PRRSV-1), PRRSV North American strain (PRRSV-2) and PCV2 was confirmed on 48%, 86% and 79% of farms, with overall prevalences of 7.6% (95% CI: 4.8–10.3%), 12.2% (95% CI: 9.6–14.7%) and 20.3% (95% CI: 14.3–26.2%) in the HKSAR pig population based on pooling results from all pigs across all farms. PRRSV-1 and PRRSV-2 were more prevalent in younger pigs, with the highest prevalences of 32.1% (95% CI: 20.8–45.0%) and 51.5% (95% CI: 38.9–64.0%) for 8-week-old pigs. In contrast, the distribution of PCV2 prevalence across age groups appeared to be more symmetrical, with higher prevalences reported in pigs from 12 weeks old to 24 weeks old but lower prevalences in younger pigs and sows. The results of this study demonstrate that PRRSV-1, PRRSV-2 and PCV2 are widely spread across pig farms in the HKSAR, which indicates that the current farm management and control protocols should be improved. We recommend the implementation of on-farm intervention strategies combined with ongoing surveillance to reduce these viruses, and their consequences, in the HKSAR pig population. Full article

The close contact between people and their pets has generated the exchange of skin microbiota, accompanied by bacteria that present resistance to antibiotics. Staphylococcus spp., opportunistic pathogens present in the skin and mucosa of mammals, have had their importance recognized in human and veterinary medicine. The objectives of this study were to identify Staphylococcus spp. present in isolates from the nostrils of healthy humans, dogs and cats as well as to determine their phenotype of resistance to methicillin. Strain identification was performed by MALDI-TOF mass spectrometry and antimicrobial susceptibility was determined using a disk diffusion assay for 12 antibiotics. Sixty humans (veterinary and technicians), sixty dogs and sixty cats were sampled; of them, 61.6%, 56.6% and 46.6%, respectively, carried Staphylococcus spp. in their nostrils, and only two people carried two different species of Staphylococcus in the only anatomical site sampled. A methicillin-resistant phenotype was present in 48.7% of the humans, 26.5% of the dogs and 57.1% of the cats, and sampled. These results demonstrate the presence of Staphylococcus spp. strains resistant to methicillin in personnel who work in contact with animals, as well as in dogs and cats that entered the same hospital or veterinary clinic, which alerts us to the potential transfer of these strains to or between people, dogs and/or cats. Full article

For the analysis of several cellular biomarkers, blood samples are anticoagulated using different agents with different modes of action. However, for the most commonly used anticoagulants, EDTA and heparin, varying effects on blood components have been reported in different species. As little is known about the impact of anticoagulants on the immunological evaluation of camel leukocytes, the present study analyzed the leukogram, the immunophenotype, and the cell vitality of camel leukocytes separated from blood samples anticoagulated with EDTA or lithium heparin. Using flow cytometry and staining with monoclonal antibodies to several cell surface markers, the composition and immunophenotype of camel leukocytes separated from blood anticoagulated with EDTA or heparin were analyzed. In comparison to EDTA-anticoagulated blood, using lithium heparin as an anticoagulant resulted in reduced numbers of total leukocytes and reduced numbers of neutrophils, which led to a reduced neutrophil to lymphocyte ratio. The analysis of cell necrosis and apoptosis after the staining of leukocytes with the DNA-sensitive dye propidium iodide and the mitochondrial membrane potential probe JC1 revealed a higher fraction of necrotic neutrophils and higher fractions of apoptotic neutrophils and monocytes in heparin blood than in EDTA blood. In addition, monocytes from heparin blood showed higher expression levels of the cell surface markers CD14, CD163, and MHCII when compared to cells from EDTA blood. Similarly, in heparin blood, CD44 and CD172a were expressed higher on neutrophils, while CD11a was expressed higher on lymphocytes in comparison to cells from EDTA blood. The results of the current study indicate the importance of considering the type of anticoagulant when investigating the composition, vitality, and immunophenotype of camel leukocytes. Full article

Luteal angiogenesis is regulated by pro-angiogenic hormones including fibroblast growth factor 2 (FGF2) and angiopoietin 1 (Ang1), which are regulated by the adipokine leptin during development. Another adipokine, adiponectin, exhibits an inverse relationship with leptin and has been identified in the CL. Therefore, it is hypothesized that adiponectin will influence pro-angiogenic hormones in the developing porcine CL. Crossbred sows were randomly allocated to one of two days of the estrous cycle, day 5 (D5; n = 4) or day 7 (D7; n = 5) for CL collection. Tissue was processed for immunohistochemical localization of adiponectin receptor 2 (AdipoR2), gene expression of FGF2, Ang1, leptin, AdipoR2, and cell culture for adiponectin treatment. The expression of AdipoR2 tended (p = 0.09) to be higher in D7 lutea and was more prevalently localized to the cell surface of large and small luteal cells than in D5 tissue. Adiponectin influenced (p ≤ 0.05) FGF2, leptin, and AdipoR2 gene expression relative to the dose and day (D5 or D7). Collectively, the evidence supports the supposition that adiponectin influences angiogenic factors in the developing CL. Full article

The objective of this study was to investigate the effect of berberine (BBR) supplementation on productivity, antioxidant markers, and the fatty acid (FA) profile in the colostrum and milk of goats. Twenty-four primiparous Saanen goats were supplemented with 0, 1, 2, and 4 g/d (per goat) of BBR in control (CON), BBR1, BBR2, and BBR4 groups (n = 6 per group), respectively, from 21 days before expected kidding to 21 days after parturition. Blood sampling was carried out at −21, −14, −7, 0, 7, 14, and 21 d relative to delivery. Colostrum was collected within the first and second milking (d 1 of lactation), and milk was harvested weekly after kidding. Both BBR2 and BBR4 increased dry matter intake (DMI) (p ≤ 0.05) and energy balance (EB) as well as colostrum and milk production. Both BBR2 and BBR4 decreased (p ≤ 0.05) plasma levels of cholesterol, haptoglobin, and ceruloplasmin, while elevating the plasma albumin and paraoxonase (p ≤ 0.05), which may indicate that BBR mitigates inflammation during the transition period. BBR reduced (p ≤ 0.05) malondialdehyde (MDA) and increased (p ≤ 0.05) total antioxidant capacity (TAC), superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and catalase (CAT) in blood, colostrum, and milk. Concentrations of de novo fatty acid in colostrum and milk were increased (p ≤ 0.05) with both BBR2 and BBR4. Free fatty acid (FFA) concentration in colostrum and milk fat were lower (p ≤ 0.05) in BBR2 and BBR4 compared to CON. The concentration of saturated fatty acids (SFAs) in colostrum and milk fat increased (p ≤ 0.05) with BBR2 and BBR4, while unsaturated fatty acids (USFAs) decreased (p ≤ 0.05) in milk. In summary, supplementation with at least 2 g/d BBR may enhance the EB and antioxidant status of dairy goats. Full article

The overpopulation of stray dogs is a serious public health and animal welfare concern in India. Neglected zoonotic diseases such as rabies and echinococcosis are transmitted at the stray–dog human interface, particularly in low to middle-income countries. The current study was designed to estimate the stray dog populations in Punjab to enhance the implementation of animal birth and disease (for example, rabies vaccination) control programs. This is the first systematic estimation of the stray dog population using a recommended method (mark–re-sight) in Punjab, India. The study was conducted from August 2016 to November 2017 in selected villages or wards in Punjab. For the rural areas, 22 sub-districts in each district were randomly selected, then one village from each of the 22 selected sub-districts was selected (by convenience sampling). For urban areas, 3 towns (less than 100,000 human population) and 2 large cities (more than or equal to 100,000 human population) were randomly selected, followed by convenience selection of two wards from each of the 5 selected towns/cities. To estimate the dog population size, we used a modified mark–re-sight procedure and analysed counts using two methods; the Lincoln–Petersen formula with Chapman’s correction, and an application of Good–Turing theory (SuperDuplicates method; estimated per km² and per 1000 adult humans and were compared between localities (villages vs. towns), dog sex (male vs. female) and age group (young vs. adult) using linear mixed models with district as a random effect. The predicted mean (95% CI) count of the dogs per village or ward were extrapolated to estimate the number of stray dogs in Punjab based on (a) the number of villages and wards in the state; (b) the adult human population of the state and (c) the built-up area of the state. Median stray dog populations per village and per ward using the Lincoln–Petersen formula with Chapman’s correction were estimated to be 33 and 65 dogs, respectively. Higher estimates of 61 per village and 112 per ward are reported using the SuperDuplicates method. The number of males was significantly higher than the number of females and the number of adult dogs was about three times the number of young dogs. Based on different methods, estimates of the mean stray dog population in the state of Punjab ranged from 519,000 to 1,569,000. The current study revealed that there are a substantial number of stray dogs and a high number reside in rural (versus urban) areas in Punjab. The estimated stray dog numbers pose a potential public health hazard in Punjab. This impact requires assessment. The estimated stray dog numbers will help develop a dog population and rabies control program in which information about the logistics required as well as costs of implementing such programmes in Punjab can be incorporated. Full article

Acupuncture has been used extensively in domestic animal medicine to treat a variety of medical conditions and diseases as an adjunct or primary therapy. Exotic animals are becoming increasingly common as pets. Owners are expecting therapies for these non-domestic animals to be similar to those available to their other domestic pets. Additionally, zoological and aquatic facilities provide medical care for the entire lives of the animals that are housed in their facilities. Many conditions similar to those observed in domestic animals can develop in zoological species and can benefit from treatment with acupuncture. Through operant conditioning or routine medical examinations, the use of acupuncture as an adjunct therapy is becoming more common. The following paper presents a summary of the types of non-domestic animals treated, for which conditions, and how these methods are commonly used. Full article

Rumen studies have focused almost exclusively on livestock species under strictly regimented diets. This means that the ruminal condition of free-living and free-feeding wildlife remains practically unstudied. Urea nitrogen salvaging, a process by which urea is passed into the rumen, to both provide a valuable source of nitrogen for bacterial growth and to buffer the potentially harmful acidic effects of bacterial short chain fatty acids, has remained unexplored in wild ruminants, such as deer. UT-B2 transporters are the key proteins reported to facilitate the transepithelial ruminal urea transport. In this study, we investigate the expression, abundance and localisation of urea transporters in the rumen of a semi-wild fallow deer (Dama dama) population. Physical measurements confirmed that males had larger rumen than females, while adults had longer papillae than juveniles. Initial RT-PCR experiments confirmed the expression of UT-B2, while immunolocalisation studies revealed that strong UT-B staining was present in the stratum basale of deer rumen. Western blotting analysis demonstrated that a 50 kDa UT-B2 protein was significantly more abundant in adult females compared to adult males. This study confirms the presence of UT-B2 urea transporters in deer rumen and suggests that sex-related differences occur, bringing new insight into our understanding of rumen physiology. Full article

Vet-OncoNet is a replicable tripartite animal cancer database with the scientific and academic purposes of collecting data and producing evidence-based knowledge for cancer science in general. Inspired by the One Health vision, Vet-OncoNet uses business intelligence tools to optimize the process of capturing, treating, and reporting animal cancer data to a national level in three interfaces: ACR (animal cancer registry, pathology-based), COR (clinical oncology registry, vet practice-based) and RFR (risk factor registry, owner-based). The first results show that skin and mammary gland are by far the most affected systems. Mast cell tumors and complex adenoma of the mammary gland are the most frequent histologic type in dogs, while in cats they are squamous cell carcinomas, tubular adenocarcinoma of the mammary gland and lymphomas. Regarding COR, it provides valuable information on the landscape of veterinary oncology practices, therapeutics options, outcomes and owners’ drivers’ adherence towards therapies, which range from 30% up to 80% upon vet practices. Furthermore, being aware of the role of animals within the family and as possible sentinels of environmental risks to cancer in humans, the network built an interface (Pet-OncoNet) dedicated to owners and a database (RFR) that receives information regarding pets and owners’ daily habits. Full article

(1) Background: Methicillin-resistant Staphylococcus aureus (MRSA) is a zoonotic pathogen that causes endocarditis, pneumonia, and skin diseases in humans and livestock. (2) Methods: The antibacterial effect of the total flavonoid against MRSA (ATCC43300) extracted from the Agrimonia pilosa Ledeb. (A. pilosa Ledeb) was evaluated by the microdilution method. The oxidative stresses in MRSA were evaluated by the levels of intracellular hydrogen peroxide (H₂O₂), reactive oxygen species (ROS), and oxidative stress-related genes. The DNA oxidative damage was tested by the 8-hydroxy-2′-deoxyguanosine (8-OHdG) and DNA gel electrophoresis. The differentially expressed proteins were determined by the method of SDS-PAGE and NanoLC-ESI-MS/MS, while the mRNAs of differential proteins were determined by Real-Time PCR. The changes of ultra-structures in MRSA were observed by Transmission Electron Microscope (TEM). (3) Results: The minimum inhibitory concentration (MIC) of the total flavonoid against MRSA was recorded as 62.5 μg/mL. After treatment with the total flavonoid, the levels of intracellular H₂O₂ and ROS were increased and the gene expressions against oxidative stress (SodA, katA, TrxB) were decreased (p < 0.01), while the gene expression for oxidative stress (PerR) was increased (p < 0.01). The level of intracellular 8-OHdG in MRSA was increased (p < 0.01) and the DNA was damaged. The results of TEM also showed that the total flavonoid could destroy the ultra-structures in the bacteria. (4) Conclusions: The total flavonoid extracted from the A. pilosa Ledeb can induce the oxidative stress that disturbed the energy metabolism and protein synthesis in MRSA. Full article

Medial compartment disease is a common occurrence in dogs affected by elbow dysplasia. Despite many treatments suggested in the literature, only few studies reported comparative outcomes in the short and long term. The aim of this study is to report and compare short- and long-term clinical and radiographic outcomes of dogs treated for medial compartment disease (MCD) by distal dynamic ulnar ostectomy (DUO), bi-oblique dynamic proximal ulnar osteotomy (BODPUO) and conservative management (CM). From 2016 to 2018, all medium to large dogs, aged between 5 and 12 months, affected by uni/bilateral MCD and treated by DUO, BODPUO or CM, were enrolled in this study and followed up for 24 months. Orthopedic and radiographic examinations were performed at T₀, T₂, T₁₂ and T₂₄ months after treatment. Lameness score, elbow arthralgia, elbow range of motion (ROM), osteoarthritis (OA) score and percentage of ulnar subtrochlear sclerosis (%STS) were evaluated at each time point. According to the treatment performed, dogs were divided into three groups: DUO, BODPUO and CM. Forty-five elbows from twenty-six dogs, treated with DUO (n = 17), BODPUO (n = 17) or CM (n = 11), were prospectively enrolled in the study. The patients enrolled in the CM group were older and showed more severe radiographic signs of OA, compared to those enrolled in the other two groups. Lameness and arthralgia scores (p < 10⁻⁴) were significantly decreased in patients that underwent surgical treatment and increased in patients managed conservatively (lameness p < 10⁻⁴, arthralgia p = 0.3068), at T₁₂ and T₂₄. OA score (p < 0.0040) and ROM (DUO, CM p < 10⁻⁴; BODPUO p = 0.0740) worsened in every study group, but %STS decreased in DUO (p = 0.0108), increased in the CM group (p = 0.0025) and remained unchanged in the BODPUO group (p = 0.2740). This study supports the clinical efficacy of DUO and BODPUO in reducing lameness, arthralgia and progression of %STS. Early diagnosis and surgical attention in patients affected by MCD can improve the short- and long-term outcome and reduce the progression of secondary changes. Full article

Similar to skin, epithelia in the tympanic membrane (TM) regenerate and move toward the opening of the external ear canal, a process called epithelial migration (EM). EM is important for maintaining healthy ears because this process removes cerumen and debris. Therefore, increasing the rate of EM or TM regeneration could be very important for healthy ear maintenance and function. Stem cells or their conditioned media have been used in medical therapy in humans to increase the rate and efficacy of EM. The purpose of this study was to evaluate the ability of canine stem cell conditioned media to accelerate EM in canine TMs. Canine adipose tissue derived-mesenchymal stem cell conditioned media (cAD-MSCCM), and several cytokines related to keratinocyte growth or migration within the media were quantified using ELISA. Ink drops were placed on the TMs of four normal beagles. Then, cAD-MSCCM was applied weekly, a total of three times to the TMs of one ear, and nothing was applied to the other eye. The results showed a higher TM EM rate in the treatment group than in the control group (p < 0.05). No adverse events were recorded. These results suggest that the weekly application of cAD-MSCCM accelerates the TM EM rate. Full article

Genetic diversity and demographic parameters were computed to evaluate the historic effects of coat colour segregation in the process of configuration of the Hispano-Arabian horse (Há). Pedigree records from 207,100 individuals born between 1884 and 2019 were used. Although coat colour is not a determinant for the admission of Hispano-Arabian individuals as apt for breeding, it may provide a representative visual insight into the gene contribution of Spanish Purebred horses (PRE), given many of the dilution genes described in Há are not present in the Arabian Purebred breed (PRá). The lack of consideration of coat colour inheritance patterns by the entities in charge of individual registration and the dodging behaviour of breeders towards the historic banning policies, may have acted as a buffer for diversity loss (lower than 8%). Inbreeding levels ranged from 1.81% in smokey cream horses to 8.80 for white horses. Contextually, crossbred breeding may increase the likelihood for double dilute combinations to occur as denoted by the increased number of Há horses displaying Pearl coats (53 Há against 3 PRE and 0 PRá). Bans against certain coat colours and patterns may have prevented an appropriate registration of genealogical information from the 4th generation onwards for decades. This may have brought about the elongation of generation intervals. Breeder tastes may have returned to the formerly officially-recognised coat colours (Grey and Bay) and Chestnut/Sorrel. However, coat colour conditioning effects must be evaluated timely for relatively short specific periods, as these may describe cyclic patterns already described in owners’ and breeders’ tastes over the centuries. Full article

Background: Endometritis is a common reproductive disease in equine animals. No investigation about the bacterial characteristics and antimicrobial susceptibility pattern of donkeys with endometritis has thus far been reported. Objectives: To determine the common uterine bacterial isolates from donkeys with endometritis and to evaluate their susceptibility to antimicrobials used for the treatment thereof. Study design: Retrospective case-series. Methods: Medical records at an equine clinical diagnostic center were retrospectively reviewed to identify submissions from donkeys with bacterial endometritis between 2018 and 2021. Data were extracted and analyzed descriptively in terms of the frequency of bacterial species, susceptibility to antimicrobials and multidrug resistance. Results: A total of 73 isolates were identified from 30 donkeys, of which 92% of the isolates were Gram-negative bacteria. Mixed cultures were found in 90% of the donkeys. The most common isolates were Escherichiacoli (31.5%) and Acinetobacter spp. (21.9%). Susceptibility testing revealed that amikacin (98%), cefoxitin (95%), trimethoprim-sulfamethoxazole (78%) and gentamicin (74%) were the most efficient agents for donkeys. Multidrug resistance (MDR) was found in 20% of all bacterial isolates, of which all Pseudomonas aeruginosa isolates showed a multidrug resistance profile. Main limitations: The sample size was relatively small, which means a bias of selection may exist. The antimicrobial resistance and MDR of agents without break points were not calculated, which means the relative results may be underestimated in our study. Conclusions: Severe infections were detected in donkeys with endometritis. Antimicrobial resistance and MDR bacteria are not rare in our study. This study demonstrated that bacteria identification and antimicrobial susceptibility testing are highly recommended before the treatment of uterine infections in donkeys. Further studies, including the epidemiological investigation of bacterial endometritis of donkeys, should be conducted to provide a better understanding of this critical problem. Full article

Heat stress is becoming a significant problem in dairy farming, especially in tropical countries, making accurate genetic selection for heat tolerance a priority. This study investigated the effect of heat stress manifestation on genetics for milk yield, milk quality, and dairy health traits with and without genomic information using single-step genomic best linear unbiased prediction (ssGBLUP) and BLUP in Thai−Holstein crossbred cows. The dataset contained 104,150 test-day records from the first lactation of 15,380 Thai−Holstein crossbred cows. A multiple-trait random regression test-day model on a temperature−humidity index (THI) function was used to estimate the genetic parameters and genetic values. Heat stress started at a THI of 76, and the heritability estimates ranged from moderate to low. The genetic correlation between those traits and heat stress in both BLUP methods was negative. The accuracy of genomic predictions in the ssGBLUP method was higher than the BLUP method. In conclusion, heat stress negatively impacted milk production, increased the somatic cell score, and disrupted the energy balance. Therefore, in dairy cattle genetic improvement programs, heat tolerance is an important trait. The new genetic evaluation method (ssGBLUP) should replace the traditional method (BLUP) for more accurate genetic selection. Full article