Vet. Sci., Volume 10, Issue 10 (October 2023) – 41 articles

Milk production continues to be the main source of income for dairy producers, and mastitis continues to be the major health challenge for dairy cows worldwide. The objective of the current study was to describe the different management practices implemented in Egyptian dairies, which may influence mastitis and improve milk quality. An in-person survey was completed with herd managers and owners of 20 Egyptian dairies selected using a stratified random sample from four of Egypt’s milk sheds. The questionnaire included 80 questions that inquired about herd demographics, mastitis status and control, milking practices, and management practices of the study dairies. Statistical analyses included descriptive statistics, multiple factor analysis (MFA), and hierarchical clustering to identify the important principal components and different dairy clusters. Of the surveyed herds, 69.50% had less than 500 lactating cows, with Holstein as the main cow breed. The reported average milk production on the study herds was 31.1 (SE ± 1.10) Kg/cow/day. Housing of lactating, dry, and close-up cows was in open lots. The majority of the dairies milked cows were three times per day (90.63%), the remaining milked cows four (5.00%) or a mix of four and three times per day. Furthermore, herringbone parlor design was the most common parlor design (66.79%) in the study dairies. The most common disinfectants used for both pre- and post-milking teat dip were iodine-based, 90.0% and 95.0%, respectively, while 52.16% of the dairies reported that their milkers wore gloves during milking. The reported mean annual percentage of mastitis was 52.3%, as a percent of the milking herd. The study dairies reported the inspection of udder health through either visual inspection (95.00%), udder hygiene score (86.88%), teat end score (71.88%), and/or using CMT (54.91%). Contagious mastitis pathogens were reported in 45.2% of the study herds. More than 50% of the study herds relied on importing pregnant cows or pregnant heifers as replacements. Multiple factor analysis identified 20 questions and represented 5 components of variability related to mastitis on dairies. The current survey of Egyptian dairies described the herd demographics and different management practices related to mastitis control and prevention. All the study dairies relied on the blanket intramammary antimicrobial drugs at dry off when current research elsewhere has identified selective dry cow therapy as an integral component of antimicrobial stewardship on dairies. Further research is required to identify the association between different management factors and the occurrence of mastitis. Full article

Argasid ticks have the vectorial potential for transmitting disease-causing pathogens to avian hosts, resulting in economic losses that may not be fully estimated. Borrelia species are the responsible agents of borreliosis in poultry, animals and humans. Our previous studies have reported a high prevalence of Argas persicus infesting domestic fowls in Khyber Pakhtunkhwa (KP), Pakistan. However, molecular screening and genetic characterization of Borrelia spp. in A. persicus have been neglected in Pakistan. In this study, we focused on the molecular epidemiology and genetic characterization of Borrelia spp. associated with A. persicus ticks infesting domestic fowls and ducks, and Carios vespertilionis infesting bats in selected districts of KP. Overall, 1818 ticks, including females (415; 23%), males (345; 19%), nymphs (475; 26%) and larvae (583; 32%), were collected from 27 locations in nine districts (Peshawar, Mardan, Swabi, Charsadda, Chitral, Lakki Marwat, Bannu, Bajaur and Hangu) from domestic fowls, ducks and their shelters, and bats. A subset of 197 ticks was selected for DNA extraction and PCR to amplify fragments of the cytochrome c oxidase (cox) gene for ticks and flagellin B (flaB) for the detection and genetic characterization of associated Borrelia spp. Among these, only Borrelia anserina DNA was detected in 40 ticks (27.2%) of different life stages, where highest prevalence was found in female ticks (18; 45%), followed by nymphs (12; 30%), larvae (7; 17.5%) and males (3; 7.5%). Tick infestation in shelters (1081; 77%) was higher than on hosts (323; 23%). The resultant cox amplicons of A. persicus showed 100% identity with the same species reported from Pakistan, China, Iran, Kenya, Kazakhstan, Algeria and Egypt and C. vespertilionis show 100% identity with the species reported from Pakistan, China, Japan, Kenya, Vietnam, Spain, Netherlands, the United Kingdom and Hungry, and clustered with the aforementioned species in the phylogenetic tree. The obtained Borrelia sequences showed 100% identity with B. anserina and revealed a close resemblance to the relapsing fever group and clustered in a monophyletic clade with B. anserina from India, Iran and Brazil in a phylogenetic tree. These results establish the first molecular characterization of B. anserina in A. persicus infesting domestic fowls and ducks in the region, as well as their shelters. To effectively control zoonotic consequences, country-wide surveillance research should be encouraged to screen soft ticks infesting various birds for associated pathogens. Full article

We sought to document the contemporary feline euthanasia practices among Australian veterinarians and determine the factors associated with the administration of a premedication or sedation prior to euthanasia. Australian veterinarians who had euthanised at least one cat during the previous 12 months were invited to participate in an anonymous online survey. From 615 valid responses, 585 respondents (95.1%) had euthanised at least one cat in the last 12 months, of which 462 respondents (75.1%) had performed an emergency euthanasia. Intravenous (IV) injection (n = 536, 91.6%) of pentobarbitone sodium (n = 584, 99.8%) was the predominant primary method. Premedication or sedation was administered to cats by 415/585 (71.0%) and 242/462 (52.4%) of respondents in non-emergency and emergency euthanasia, respectively. In a multivariable logistic regression analysis, for non-emergency euthanasia, being female and working in a metropolitan area were significant predictors for administering a premedication or sedation (p < 0.001 and p = 0.037, respectively). For emergency euthanasia, working in an ‘other’ type of practice was a significant predictor for administering a premedication or sedation (p < 0.001). Australian veterinarians vary in their approach to feline euthanasia. There is scope for refinement of euthanasia techniques to maximise the welfare of cats, their owners or guardians, and veterinary team members. Full article

The present case describes multiple hepatic lipomas in a common hill mynah (Gracula religiosa). A 21-year-old female captive common hill mynah died without any notable clinical symptoms. An autopsy and histopathological examinations were conducted to determine the exact cause of death. On gross observation, the liver demonstrated a yellowish white surface color and multiple nodules indicating neoplastic lesions. Histopathological assessment found that the nodules purely comprised mature adipocytes. Furthermore, the liver exhibited an excessive accumulation of iron in hepatocytes and Kupffer cells and the infiltration of chronic inflammatory cells, suggesting iron storage disease (ISD). Based on the results, the present case was diagnosed as multiple hepatic lipomas with ISD. To the authors’ best knowledge, multiple hepatic lipomas accompanying ISD lesions have not been reported previously. Hence, the present case is the first case report of hepatic multiple lipomas with ISD in veterinary medicine. Full article

Lumpy skin disease virus (LSDV) infection, accompanied by loss of hide quality, poor reproductive efficiency, consistent degenerative emaciation, and milk yield reduction of animals, causes severe economic implications in endemic zones. The heterologous attenuated goat pox (GTPV) vaccine (AV41 strain) was used in China to prevent LSDV infection. Only a few LSDV detection methods that distinguish LSDV from GTPV vaccine strains have been reported before. For simple, rapid, and specific detection of LSDV, the real-time recombinase polymerase amplification (RPA) method was established with the specific primers and probes designed according to the conserved regions of ORF132 gene sequences. The assay could be finished within 20 min at a constant temperature (39 °C). This method had a limit of detection (LOD) of 15 copies/μL for LSDV and no cross-reaction with the nucleic acids of goat pox virus, infectious bovine rhinotracheitis virus, Pasteurella multocida, and bovine healthy tissue. Furthermore, 43 clinical samples were detected by this method and the real-time PCR recommended by the World Organisation for Animal Health (WOAH), with a kappa value, was 0.94. These results demonstrated that the real-time RPA method for detecting LSDV developed in this study was characterized by high sensitivity and specificity, which has wide application value in the clinical diagnosis and detection of LSDV in China. Full article

The purpose of the present research was to define ovarian follicular dynamics and plasma endocrine profiles in response to a single PGF_2α injection, administered indiscriminately during the breeding season of Barbari goats. Ovarian dynamics were observed at every 12 h interval by using B mode ultrasonography, blood samples for hormonal analysis such as estradiol 17β and progesterone were collected at every 12 h interval, and bucks with aprons were used to identify standing estrus at every 6 h interval. Relative to PGF_2α, the start of standing estrus and ovulation differ (p < 0.05) between early- (n = 7), intermediate- (n = 6), and late-responding (n = 6) goats. The highest plasma level of estradiol 17β was detected 12 h prior to ovulation. The average diameter of the ovulatory follicle and length of standing estrus were comparable (p > 0.05) between the goats. The corpus luteum degenerated more quickly (p < 0.05) in early- than intermediate- and late-responding goats. Dominant follicle diameter and estradiol 17β concentration also differ (p < 0.05) among groups. Although the plasma level of progesterone did not vary (p = 0.065), the variation in progesterone concentration with time differed (p < 0.05) amongst the goats. As a result, this research indirectly reveals that the beginning of standing estrus, end of estrus, and ovulation after PGF_2α might fluctuate in Barbari goats because of follicular and hormonal dynamics during the luteal phase. Full article

Ceruloplasmin (Cp) assessment in biological samples exploits the oxidase activity of this enzyme against several substrates, such as p-phenylenediamine (p-P), o-dianisidine (o-D) and, most recently, ammonium iron(II) sulfate (AIS). Once developed in humans, these assays are often used in veterinary medicine without appropriately optimizing in the animal species of interest. In this study, two assays using AIS and o-D as substrates have been compared and validated for Cp oxidase activity assessment in horse’s plasma. The optimization of the assays was performed mainly by varying the buffer pH as well as the buffer and the substrate molar concentration. Under the best analytical conditions obtained, the horse blood serum samples were treated with sodium azide, a potent Cp inhibitor. In the o-D assay, 500 µM sodium azide treatment completely inhibits the enzymatic activity of Cp, whereas, using the AIS assay, a residual analytical signal was still present even at the highest (2000 µM) sodium azide concentration. Even though the analytical values obtained from these methods are well correlated, the enzymatic activity values significantly differ when expressed in Units L⁻¹. A disagreement between these assays has also been detected with the Bland–Altman plot, showing a progressive discrepancy between methods with increasing analytical values. Full article

Rabbit production holds significant relevance in modern agriculture due to its potential as a sustainable source of high-quality protein and efficient feed conversion, contributing to food security and economic diversification. Nevertheless, studies incorporating feto-maternal monitoring in this species are uncommon. This review gathers research on the monitoring and evaluation of factors affecting rabbit gestation, providing a better understanding of the causes of prenatal development abnormalities. These include studies regarding how chronic maternal hypertension, gestational diabetes, maternal stress, ectopic gestation, maternal uterine ischemia and fetal hypoxia, intrauterine growth restriction, superfetation, maternal age, maternal nutritional status, maternal physical condition, maternal and embryonic genotype, and the intrauterine location of rabbit fetuses can potentially impact rabbits’ reproduction and maternal and fetal health. Among other monitoring techniques, ultrasonography, considered one of the best tools for diagnosing pregnancy and conducting follow-up, is also reviewed. Details on measurable fetal-development parameters in rabbits and precautions to be considered before and during the examination are also provided. Additional studies are required to understand why some events occur and their consequences throughout gestation, allowing the determination of new biomarkers or cut-offs that can be helpful for early diagnosis and improve reproductive efficiency. Full article

Biological variation (BV) describes the physiological random fluctuation around a homeostatic set point, which is a characteristic of all blood measurands (analytes). That variation may impact the clinical relevance of the changes that are observed in the serial results for an individual. Biological variation is represented mathematically by the coefficient of variation (CV) and occurs within each individual (CV_I) and between individuals in a population (CV_G). Biological variation data can be used to assess whether population-based reference or subject-based reference intervals should be used for the interpretation of laboratory results through the calculation of the index of individuality (IoI). This study aimed to determine the biological variations, calculate the IoI and reference change values (RCV) of clinical chemistry analytes in an outbred strain colony of Hartley guinea pigs (GPs), and set the quality specifications for clinical chemistry analytes. Blood was collected from 16 healthy adult laboratory colony GPs via jugular venipuncture at weekly intervals over six weeks. All the samples were frozen and analyzed in a single run. Analytical, CV_I, and CV_G biological variations, together with the IoI and RCV, were calculated for each measurand. Based on the estimated BV, the calculated IoI was low for glucose, so individual reference intervals (RCV) should be used. The majority of the measurands should be interpreted using both population-based and subject-based reference intervals as the IoIs were intermediate. Full article

Measuring C-reactive protein (CRP) in serum is a useful surrogate marker for assessing disease progression and treatment response in dogs with autoinflammatory diseases. Affected dogs often receive high-dose glucocorticoid treatment, but the effect of such treatment alone on serum CRP concentrations is unknown. We evaluated serum CRP concentrations via immunoassay (sandwich enzyme-linked immunosorbent assay and particle-enhanced turbidimetric immunoassay) in 12 healthy beagle dogs administered high-dose hydrocortisone (8 mg/kg q12 h) per os vs. placebo over 28 days (days 0, 1, 5, and 28) in a randomized parallel study design. Serum CRP concentrations slightly decreased during treatment or placebo but without a significant association with hydrocortisone administration (p = 0.761). Compared to baseline, serum CRP concentrations were decreased by >2.7-fold (minimum critical difference) in three hydrocortisone-treated dogs and two dogs in the placebo group on day 28, whereas an increase to >2.7-fold was seen in one dog receiving placebo. These results suggest a lack of confounding effects of high-dose hydrocortisone administration on serum CRP concentrations in healthy dogs. This might also hold in dogs with autoinflammatory conditions and/or administration of other high-dose corticosteroids, suggesting that CRP presents a suitable biomarker to monitor inflammatory disease processes. However, this needs confirmation by further studies evaluating corticosteroid-induced cellular (e.g., hepatic) transcriptome and proteome changes. Full article

Palatine tonsils are lymphoid organs, whose anatomic localization gives them a role against antigens entering the body during feeding and breathing. In human medicine, MRI is used to investigate tonsillar diseases. In veterinary medicine, a recent study on healthy dogs described the MRI appearance of canine palatine tonsils, with no available reports about feline ones. Due to the similarities between animals and humans, and based on the study on canine tonsils, the authors aimed to evaluate the feasibility of low-field MRI to detect and describe presumed normal features of feline palatine tonsils, assessing the finding’s reproducibility. Low-field MRI of the heads of 14 cats was reviewed, and qualitative findings (visualization, shape, margins, signal intensity, and pattern) and size of each tonsil were recorded. Each observer recorded 71% of the expected tonsils. Most of them were classified as oval, ill-defined, and hyperintense structures with both homogeneous and heterogeneous signal patterns; the overall agreement was considered good. Low-field MRI is potentially a useful imaging modality to visualize palatine tonsils in cats, and their normal appearance has been described for the first time. The authors recommend the evaluation of tonsils in the transverse plane and consider the most accurate estimation of the short axis. Full article

Nutrition and health during pre-weaning affect the calves’ future fertility, calving age, production, and carrier length. Calves are highly susceptible to neonatal calf diarrhea (NCD), which can be fatal. NCD is due to hypovolemia and acidosis, which may involve anorexia and ataxia. The One Health principle calls for a drastic reduction in antimicrobial use. One approach is to improve animal health and reduce the use of antibiotics and functional ingredients that have beneficial effects due to bioactive compounds. Several functional ingredients and additives can be considered, and, in particular for this study, Ascophyllum nodosum was considered. The present study aimed to evaluate the role of A. nodosum as a functional ingredient implemented into the milk replacer in neonatal calves. Twelve pre-weaned Holstein Frisian calves, housed in twelve individual pens in the same environmental conditions, were divided into two groups of six animals: a control group (CTRL, n = 6) fed with a milk replacer, and a treatment group receiving milk enriched with 10 g of A. nodosum in their diet (TRT, n = 6) for 42 days. The fecal score was evaluated daily (3–0 scale) to monitor the incidence of diarrhea in the two groups. The body weight was evaluated weekly, and every two weeks feces were collected for microbiological evaluation using a selective medium for plate counting of total, lactic acid, and coliform bacteria. To verify the presence of Lactobacillus, Bifidobacterium, and Escherichia coli, real-time qPCR was used. At the beginning and at the end of the trial, blood samples were obtained for serum metabolite analysis. The growth performance did not differ in either of the two groups, but significant differences were observed in the incidence of moderate diarrhea (p-value < 0.0113), where the TRT group showed a lower incidence of cases during the 42-day period. Serum analysis highlighted higher contents of albumin, calcium, phosphorus, and total cholesterol in the TRT group compared to CTRL (p-value < 0.05). In conclusion, implementation of A. nodosum in the diet of calves can lead to better animal welfare and may reduce the use of antibiotics. Full article

This study aimed to evaluate the effectiveness of different additives in preventing urolithiasis in lambs fed a diet rich in concentrate and their impact on performance and blood and urinary parameters. Thirty-two noncastrated male lambs, crossbred Dorper × Santa Inês, with initial body weights (BWs) of 23 ± 0.1 kg and ages of 50 ± 5 days, were kept in individual pens and fed a diet composed of 6% Cynodon ssp. hay and 94% concentrate and subjected to four treatments: CON without inclusion of additives, addition of ACL 5 g/kg of dry matter (DM), addition of CCL 6.3 g/kg of DM, and addition of BZA 5 g/kg of DM. There was no effect of treatment or interaction with time on blood parameters (p > 0.050), and performance characteristics, morphometry of ruminal papillae, and scores of cecum and rumen lesions were not affected by the addition of additives to the diet (p > 0.050). Greater urinary acidification was observed in animals from CCL and ACL treatments (p = 0.033). Calcium chloride acidified the urinary pH and can be used instead of ammonium chloride at a concentration of 0.63% based on DM, when this is the objective. Full article

Bovine mastitis is an inflammation of the udder tissue of the mammary gland brought on by microbial infections or physical damage. It is characterised by physical, chemical, and biological changes in the udder and milk. While several different bacterial species have been identified as causative agents of mastitis, many subclinical mastitis (SCM) cases remain culture-negative. The aim of this study was to characterise milk microbiota from SCM and apparently healthy dairy cows (non-SCM) by 16S rRNA sequencing. Alpha-diversity metrics showed significant differences between SCM cows and non-SCM counterparts. The beta-diversity metrics in the principal coordinate analysis significantly clustered samples by type (PERMANOVA test, p < 0.05), while non-metric dimensional scaling did not (PERMANOVA test, p = 0.07). The overall analysis indicated a total of 95 phyla, 33 classes, 82 orders, 124 families, 202 genera, and 119 bacterial species. Four phyla, namely Actinobacteriota, Bacteroidota, Firmicutes, and Proteobacteria collectively accounted for more than 97% of all sequencing reads from SCM and non-SCM cow samples. The most abundant bacterial classes were Actinobacteria, Bacilli, Bacteroidia, Clostridia, and Gammaproteobacteria in non-SCM cow samples, whilst SCM cow samples were mainly composed of Actinobacteria, Alphaproteobacteria, Bacilli, Clostridia, and Gammaproteobacteria. Dominant bacterial species in non-SCM cow samples were Anthropi spp., Pseudomonas azotoformans, P. fragi, Acinetobacter guillouiae, Enterococcus italicus, Lactococcus lactis, whilst P. azotoformans, Mycobacterium bovis, P. fragi, Acinetobacter guillouiae, and P. koreensis were dominant in the SCM cow samples. The current study found differences in bacterial species between SCM and non-SCM cow milk; hence, the need for detailed epidemiological studies. Full article

To evaluate the acute and chronic 28-day repeated-dose oral toxicity of Guixiong Yimu San (GYS) in mice and rats, high-performance liquid chromatography (HPLC) was used to determine the stachydrine hydrochloride in GYS as the quality control. In the acute toxicity trial, the mice were administered orally at a dose rate of 30.0 g GYS/kg body weight (BW) three times a day. The general behavior, side effects, and death rate were noticed for 14 days following treatment. In the subacute toxicity trial, the rats were administered orally at a dose rates of30.0, 15.0, and 7.5 g GYS/kg BW once a day for 28 days. The rats were monitored every day for clinical signs and deaths; changes in body weight and relative organ weights (ROW) were recorded every week, hematological, biochemical, and pathological parameters were also examined at the end of treatment. The results showed that the level of stachydrine hydrochloride in GYS was 2.272 mg/g. In the acute toxicity trial, the maximum-tolerated dose of GYS was more than 90.0 g/kg BW, and no adverse effects or mortalities were noticed during the 14 days in the mice. At the given dose, there were no death or toxicity signs all through the 28-day subacute toxicity trial.The oral administration of GYS at a dose rate of 30.0 g/kg/day BW had no substantial effects on BW, ROW, blood hematology, gross pathology, histopathology, and biochemistry (except glucose), so 30.0 g/kg BW/day was determined as the no-observed-adverse-effect dosage. Full article

We measured the levels of prednisolone (PSL) residues in milk of intramuscularly dosed dairy cows and established a withdrawal time (WT) of PSL in milk. Eight healthy Holstein cows were injected with 10 (PSL-1) or 20 (PSL-2) mL of 10 mg/mL of PSL, and then, their milk was sampled at 12 h intervals for five days. PSL residue concentrations in milk were determined using LC-MS/MS. The correlation coefficient of the calibration curve was 0.9976. The limit of detection (LOD) and the limit of quantification (LOQ) were 0.2 µg/kg and 0.6 μg/kg, respectively. Recoveries ranged from 96.5% to 110.0%, and the coefficient of variation was <5.64%. At 24 h after administration, PSL levels in PSL-1 and PSL-2 were below the LOQ in all milk samples. Although this study had a smaller sample size than the European Medicines Agency’s recommendations (n = 20), it was based on the Animal and Plant Quarantine Agency guidelines of the Republic of Korea (n = 8) for the determination of withdrawal periods in milk. We established the withdrawal period for both PSL-1 and PSL-2 in milk at 12 h. In conclusion, we developed an analytical method that is sensitive and can reliably detect PSL in milk, and our estimated WT of PSL in bovine milk is shorter than the current 3-day withdrawal period of PSL in commercial PSL products. Full article

Allergen-specific immunotherapy (AIT) is the only current intervention that has the ability to modify the immune response toward a tolerogenic state. This study aimed to assess the efficacy and safety of AIT in horses with allergic diseases in a systematic manner. Three databases were searched to identify articles reporting clinical outcomes and adverse events associated with AIT. The articles were evaluated for beneficial responses to AIT, defined as a ≥50% reduction in clinical signs, and clinical remission. Horses with respiratory diseases, urticaria, and pruritic dermatitis receiving insect monotherapy or multi-allergen AIT were included. All adverse events were graded, and analytical and confounding biases were assessed. The results showed that multi-allergen AIT had a beneficial response in 75% of horses with respiratory diseases, 88% with urticaria, and 56% with pruritic dermatitis. However, horses treated solely with insect AIT for pruritic dermatitis had a lower response rate (36%). Self-limiting local reactions were the most common adverse events, with systemic reactions grade II accounting for 11% of reported events. Analytical and confounding biases were identified as major limitations in the available studies. Further research is needed to address these biases and provide stronger evidence on the efficacy and safety of AIT in horses with allergic diseases. Full article

Phytogenic compounds can improve feed efficiency, meat quality, and the health status of chickens under hot climatic conditions. The current study investigated the impact of the bioactive lipid compounds of oregano and peppermint and their combination on the sustainability of meat production and the health of broiler chicks in hot climatic conditions. Two hundred and fifty-six one-day-old broiler chicks were distributed into four treatment groups. The birds were fed a control diet, bioactive lipid compounds of oregano (BLCO, 150 mg/kg), bioactive lipid compounds of peppermint (BLCP, 150 mg/kg), or a combination of BLCO and BLCP at 150 mg/kg each for 35 days. Each treatment included 8 replicates, each with 8 birds. The results showed that adding BLCO and BLCP separately or in combination to broiler diets improved body weight, body weight gain, and feed conversion ratio. BLCO, BLCP, or their combination increased the percentages of the dressing and gizzard and lowered the percentage of abdominal fat as compared to the control. Supplementation of BLCO, BLCP, or their combination decreased serum cholesterol, triglycerides, aspartate aminotransferase, alanine transaminase, creatinine, and urea compared to control. BLCO, BLCP, or their combination reduced cook and drip loss in the meat of broilers. In conclusion, birds fed diets containing BLCO and BLCP, either independently or in combination, showed improvements in performance, blood biochemistry, and meat quality in hot climatic conditions. Full article

Mature cataracts can be a life-threatening condition in fish as it may result in anorexia. Lens extraction has been previously described in fish, but the long-term outcome of this procedure has not been evaluated. Eleven captive-bred adult fish housed in a public aquarium presented with unilateral (n = 3/11) or bilateral (n = 8/11) mature cataracts. All cases belonged to three endangered fish populations: striped bass (Morone saxatilis) originating from the Saint Lawrence River and Atlantic and spotted wolffish (Anarhichas lupus and Anarhichas minor). Pre-operatively, fish presented with anorexia, dark discoloration, abnormal position in the water column, skin abrasions, and/or decreased body condition scores. A lensectomy was performed in eleven fish, including a bilateral procedure in four fish, corresponding to fifteen eyes. Follow-up examinations were performed one to two weeks and one year post surgery. The main complication was self-resolving: corneal edema was present in 67% of eyes, persistent after a week in 47% of eyes, and resolved thereafter. Post-operatively, 73% of fish resumed feeding and regained a good body condition score. Persistent visual impairment post-surgery associated with anorexia required euthanasia in three out of eleven cases. The median time of death was 336 days post surgery (range: 27–1439 days) and three cases were still alive 1334 to 1425 days after the lensectomy. This was considered a favorable outcome for these older individuals with concurrent diseases. Full article

This study aimed to evaluate the proteomic profile of seminal plasma from young Nellore bulls. We used 20 bulls aged between 19.8 and 22.7 months, divided into two groups according to the results of the Breeding Soundness Evaluation (BSE): approved (FIT n = 10) and not approved (UNFIT n = 10). The scrotal perimeter was measured and a semen collection was performed through electroejaculation. The percentage of sperm motility, mass motility, and sperm vigor were calculated using conventional microscopy, and the percentage of sperm abnormalities was calculated using phase-contrast microscopy of all ejaculates. Seminal plasma was separated from spermatozoa using centrifugation and processed for proteomic analysis by LC-MS/MS. Seminal plasma proteins were identified using MASCOT Daemon software v.2.4.0 and label-free quantification analysis was carried out by SCAFFOLD Q+ software v.4.0 using the Exponentially Modified Protein Abundance Index (emPAI) method. Functional classification of proteins was performed based on their genetic ontology terms using KOG. Functional cluster analysis was performed on DAVID. There were no differences in scrotal perimeter and physical semen characteristics between FIT and UNFIT groups of bulls. The percentage of sperm abnormalities was higher (p < 0.05) in the UNFIT group of bulls. A total of 297 proteins were identified for the two groups. There were a total of 11 differentially abundant proteins (p < 0.05), two of them more abundant in FIT bulls (Spermadhesin-1 and Ig gamma-1 chain C region) and nine in UNFIT bulls (Vasoactive intestinal peptide, Metalloproteinase inhibitor 2, Ig lambda-1 chain C regions, Protein FAM3C, Hemoglobin beta, Seminal ribonuclease, Spermadhesin 2, Seminal plasma protein BSP-30kDa, and Spermadhesin Z13). Spermadhesin-1 was the protein with the highest relative abundance (36.7%) in the seminal plasma among all bulls, corresponding to 47.7% for the FIT bulls and 25,7% for the UNFIT bulls. Posttranslational modification, protein turnover, and chaperones were the functional categories with the highest number of classified proteins. Protein functional annotation clusters were related to Phospholipid efflux, ATP binding, and chaperonin-containing T-complex. The differentially abundant proteins in the group of FIT bulls were related to sperm capacitation and protection against reactive species of oxygen. In contrast, differentially expressed proteins in the group of UNFIT bulls were related to motility inhibition, intramembrane cholesterol removal and oxidative stress. In conclusion, the proteomic profile of the seminal plasma of FIT bulls presents proteins with participation in several biological processes favorable to fertilization, while the proteins of the seminal plasma of UNFIT bulls indicate a series of alterations that can compromise the fertilizing capacity of the spermatozoa. In addition, the relative abundance of spermadhesin-1 found in the seminal plasma of young Nellore bulls could be studied as a reproductive parameter for selection. Full article

Molecular diagnostic tests have evolved very rapidly in the field of human health, especially with the arrival of the recent pandemic caused by the SARS-CoV-2 virus. However, the animal sector is constantly neglected, even though accurate detection by molecular tools could represent economic advantages by preventing the spread of viruses. In this regard, the swine industry is of great interest. The main viruses that affect the swine industry are described in this review, including African swine fever virus (ASFV), porcine reproductive and respiratory syndrome virus (PRRSV), porcine epidemic diarrhea virus (PEDV), and porcine circovirus (PCV), which have been effectively detected by different molecular tools in recent times. Here, we describe the rationale of molecular techniques such as multiplex PCR, isothermal methods (LAMP, NASBA, RPA, and PSR) and novel methods such as CRISPR-Cas and microfluidics platforms. Successful molecular diagnostic developments are presented by highlighting their most important findings. Finally, we describe the barriers that hinder the large-scale development of affordable, accessible, rapid, and easy-to-use molecular diagnostic tests. The evolution of diagnostic techniques is critical to prevent the spread of viruses and the development of viral reservoirs in the swine industry that impact the possible development of future pandemics and the world economy. Full article

Dogs are considered the major domestic reservoir for human visceral leishmaniasis, a serious disease caused by the Leishmania infantum parasite. Diagnosis of canine visceral leishmaniasis (CVL) is critical for disease control, with several methods currently available. Among the serological tests, the DPP rapid test and the EIE-LVC, more commonly used in Brazil, are associated with variable sensitivity and specificity. Research with novel recombinant proteins such as the ELISA with the recombinant chimeric protein Q5 may therefore improve the CVL diagnosis. This study aimed to evaluate the true diagnostic potential of Q5 in an ELISA assay using a large number of CVL-suspected sera (406) with a previous positive diagnosis based on the rapid DPP test. Sera from the DPP-positive dogs, also assessed with the EIE-LVC test, were compared with sera from healthy dogs (n = 46) and used for ELISA tests using the recombinant Q5. The resulting data as well as the correlation with the clinical signs and the environmental characteristics of the animals were analyzed using Medal and GraphPad Prism 8.0. Overall, similar levels of lower sensitivity (67–68%) were seen for both the commercial EIE-LVC test and the Q5 ELISA when all assessed sera were considered, but a much greater sensitivity (92%) was seen for those samples from symptomatic dogs only. In contrast, many negative results were observed for the DPP-positive sera from asymptomatic dogs or those with no clinical information available. A selection of those sera were tested yet again in new ELISA assays using a second batch of the recombinant Q5, purified under milder denaturing conditions, as well as using another recombinant protein (Lci13). The results reveal a higher-than-expected incidence of likely false-positive results for DPP, reinforcing the need for other recombinant proteins, such as the chimeric Q5, to be investigated as possible alternatives to the currently used CVL diagnostic methods. Full article

Mammary cancer is one of the main causes of death in female dogs worldwide, considering that many risk factors are involved in its development. This study aimed to elucidate the relationship between epidemiological and clinical risk factors with the histopathological diagnosis of malignant mammary tumors in dogs treated at the Veterinary Hospital of the Federal University of Uberlândia, which has one of the first veterinary oncology services in Brazil. A retrospective matched case-control study was conducted to identify risk factors for the development of malignant mammary tumors in dogs. The variables analyzed were size dog, breed, housing, type of diet, and body score. Potential risk factors were selected by univariate analysis (p < 0.25) before multivariate forward binary logistic regression. The most frequent benign tumor was the benign mixed tumor (35.2%), and the most frequent malignant tumor was the mixed carcinoma (27.4%). Size dog, breed, housing, and overweight are predictors of malignant mammary tumors in dogs. The highest risk of developing malignant mammary tumors is associated with large female dogs, Yorkshire or Poodle breeds, living outside the home, or being overweight. Full article

This research aimed to explore cat and dog owners’ attitudes and opinions towards various pet care practices in Aotearoa New Zealand (NZ), and determine what factors were associated with them. An online survey composed of both closed and open-ended questions was administered from January to March 2019. A total of 2358 respondents answered the questions on pet care practices. Of these, 37.5 percent (n = 885) were both dog and cat owners, 28.0 percent (n = 652) were cat owners, and 26.0 percent (n = 609) were dog owners, while 9% (n = 212) of respondents did not own a cat or dog at the time of taking the survey. The study revealed that most respondents (>90%) acknowledge the importance of providing adequate housing, regular worming and flea treatments, microchipping, and vaccinations for their pets. Notably, demographic factors such as gender, ethnicity, age range, having children, having a rural upbringing, and place of residence influenced owners’ attitudes, with those towards cat care practices varying more than those for dogs. The study highlights the need for responsible pet ownership interventions considering these demographic factors. The study identifies a knowledge gap among pet owners concerning the importance of regular veterinary visits and the impact of breeding for certain looks on animal welfare. The use of shock collar devices provoked varying opinions on their usage for training and behavior modification. The study suggests that enhancing pet owners’ knowledge is pivotal for responsible pet ownership. Overall, the findings emphasize the need for tailored interventions that account for demographic variations for promoting responsible pet ownership and animal welfare. The findings underscore the importance of improving access to veterinary health care teams, especially in rural areas, and of providing culturally appropriate education resources for both pet owners and veterinary health care teams. Full article

Ticks are hematophagous ectoparasites that transmit pathogens to animals and humans. Updated knowledge regarding the global epidemiology of tick-borne Rickettsia hoogstraalii is dispersed, and its molecular detection and genetic characterization are missing in Pakistan. The current study objectives were to molecularly detect and genetically characterize Rickettsia species, especially R. hoogstraalii, in hard ticks infesting livestock in Pakistan, and to provide updated knowledge regarding their global epidemiology. Ticks were collected from livestock, including goats, sheep, and cattle, in six districts of Khyber Pakhtunkhwa (KP) Pakistan. Overall, 183 hosts were examined, of which 134 (73.2%), including goats (number = 39/54, 72.2%), sheep (23/40, 57.5%), and cattle (71/89, 80%) were infested by 823 ticks. The most prevalent tick species was Rhipicephalus microplus (number = 283, 34.3%), followed by Hyalomma anatolicum (223, 27.0%), Rhipicephalus turanicus (122, 14.8%), Haemaphysalis sulcata (104, 12.6%), Haemaphysalis montgomeryi (66, 8.0%), and Haemaphysalis bispinosa (25, 3.03%). A subset of 210 ticks was selected and screened for Rickettsia spp. using PCR-based amplification and subsequent sequencing of rickettsial gltA and ompB fragments. The overall occurrence rate of R. hoogstraalii was 4.3% (number = 9/210). The DNA of Rickettsia was detected in Hy. anatolicum (3/35, 8.5%) and Ha. sulcata (6/49, 12.2%). However, no rickettsial DNA was detected in Rh. microplus (35), Rh. turanicus (35), Ha. montgomeryi (42), and Ha. bispinosa (14). The gltA and ompB fragments showed 99–100% identity with R. hoogstraalii and clustered phylogenetically with the corresponding species from Pakistan, Italy, Georgia, and China. R. hoogstraalii was genetically characterized for the first time in Pakistan and Hy. anatolicum globally. Further studies should be encouraged to determine the role of ticks in the maintenance and transmission of R. hoogstraalii in different hosts. Full article

The ability of bovine oocytes to reach the blastocyst stage (i.e., embryo with around 150 cells in cattle) in vitro can be affected by technical (e.g., culture medium used) and physiological factors in oocyte donors (e.g., age, breed). As such, the nutritional status of oocyte donors plays a significant role in the efficiency of in vitro embryo production (IVEP), and several nutritional strategies have been investigated in cattle subjected to ovum pick-up (OPU). However, there is no clear consensus on the reliability of nutritional schemes to improve IVEP in cattle. Available evidence suggests that a moderate body condition score (i.e., 3 in a 1–5 scale) in cattle is compatible with a metabolic microenvironment in ovarian follicles that will promote embryo formation in vitro. The usefulness of fatty acid and micronutrient supplementation to improve IVEP in cattle is debatable with the current information available. Overall, the supply of maintenance nutritional requirements according to developmental and productive stage seems to be enough to provide bovine oocyte donors with a good chance of producing embryos in vitro. Future nutrition research in cattle using OPU-IVEP models needs to consider animal well-being aspects (i.e., stress caused by handling and sampling), which could affect the results. Full article

Rumen flukes are geographically widespread trematodes affecting wild and domestic ruminants. The juvenile forms, which are found in the small intestine, are more pathogenic compared to the adults. Severe diarrhoea and weight loss are the major clinical signs, and the disease might be fatal in severely infested individuals. In the last decade, paramphistomosis has been described as an emerging parasitic disease in Europe. This study aimed to identify the rumen fluke species in wild ruminants from western Romania. Fifty-two pre-stomachs obtained from roe deer (Capreolus capreolus) that were hunted on 14 hunting grounds from Timiș and Arad counties were examined for the presence of paramphistomes. Three (9.09%) out of 33 samples were positive in Timiș County, and one (5.26%) out of 19 samples was positive in Arad County. Subsequent PCR testing revealed that three samples were positive for Calicophoron daubneyi and one for Paramphistomum leydeni. The presence of C. daubneyi and P. leydeni in roe deer has not been previously reported in Romania. Two Paramphistomum species—C. daubneyi and P. leydeni—were revealed as the main species of rumen flukes in roe deer from forests in Romania. Full article

Tracheal hypoplasia is a major concern in brachycephalic dogs, but there is no consensus for the trachea in brachycephalic cats. We aimed to compare tracheal length and diameter between normo- and brachycephalic cats using computed tomography (CT) image measurements and evaluate their usefulness in tracheostomy planning. A total of 15 normocephalic and 14 brachycephalic cats were included in the study. Tracheas of normocephalic cats were significantly longer compared with brachycephalic cats. No difference was detected in tracheal diameter between normocephalic and brachycephalic cats. Both groups had a lateral diameter significantly larger than the dorsoventral diameter at the level of the cranial end of the manubrium sterni and at the level of the second rib. Normocephalic and brachycephalic cats’ tracheas have the same dorsoventral flattening at the level of the cranial end of the manubrium sterni and at the level of the second rib. The location between the 4th and 5th cervical vertebrae seems the best place to perform a tracheostomy in cats due to its round shape and easily accessible anatomical location. No sign of tracheal hypoplasia in brachycephalic cats was detected. Finally, 7 mm appears to be an adequate diameter for the tracheal tubes used to perform feline tracheostomies. Full article

The diagnostic workup of respiratory disease in pigs is complex due to coinfections and non-infectious causes. The detection of pathogens associated with respiratory disease is a pivotal part of the diagnostic workup for respiratory disease. We aimed to report how frequently certain viruses and bacteria were detected in samples from pigs with respiratory symptoms in the course of routine diagnostic procedures. Altogether, 1975 routine diagnostic samples from pigs in Austrian swine stocks between 2016 and 2021 were analysed. PCR was performed to detect various pathogens, including porcine reproductive and respiratory syndrome virus (PRRSV) (n = 921), influenza A virus (n = 479), porcine circovirus type 2 (PCV2) (n = 518), Mycoplasma (M.) hyopneumoniae (n = 713), Actinobacillus pleuropneumoniae (n = 198), Glaesserella (G.) parasuis (n = 165) and M. hyorhinis (n = 180). M. hyorhinis (55.1%) had the highest detection rate, followed by PCV2 (38.0%) and Streptococcus (S.) suis (30.6%). PRRSV was detected most frequently in a pool of lung, tonsil and tracheobronchial lymph node (36.2%). G. parasuis was isolated more frequently from samples taken after euthanasia compared to field samples. PRRSV-positive samples were more likely to be positive for PCV2 (p = 0.001), M. hyopneumoniae (p = 0.032) and Pasteurella multocida (p < 0.001). M. hyopneumoniae-positive samples were more likely to be positive for P. multocida (p < 0.001) and S. suis (p = 0.046), but less likely for M. hyorhinis (p = 0.004). In conclusion, our data provide evidence that lung samples that were positive for a primary pathogenic agent were more likely to be positive for a secondary pathogenic agent. Full article

The Bísaro pig is a Portuguese autochthonous breed greatly appreciated for its meat quality and is mainly reared outdoors. Immunocastration could be a solution to avoid undesirable pregnancies and boar taint in cull sows. The present study tested three immunocastration protocols (with Improvac^®) according to their reproductive cycle. The first inoculation was performed two weeks after farrowing (IM1, n = 5), at the beginning of estrus (IM2, n = 5), and one week after the end of estrus (IM3, n = 5), followed by a second administration four weeks apart. A control group (C, n = 5) was also included in the same housing conditions. The sample collection included the reproductive tract for morphometric evaluation, neck fat for the quantification of boar taint compounds, and a portion of the Longissimus thoracis et lumborum for meat quality trait assessment. The reproductive tracts from intact sows (C) were significantly heavier compared to the immunocastrated groups (p < 0.05) (1.403 kg C to 0.508 kg IM1, 0.590 kg IM2, and 0.599 kg IM3), suggesting the regression of the reproductive tract to nonstimulated conditions due to immunization against GnRH. The IM1 group exhibited significantly smaller reproductive tract measurements compared to group C for most of the evaluated segments (p < 0.05). No marked differences were observed in the meat quality traits. Therefore, immunocastration can be used in culling sows to avoid ovarian activity, and it is not detrimental to pork quality traits. Full article