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Flavobacterium psychrophilum, the etiological agent of bacterial coldwater disease and rainbow trout fry syndrome, causes great economic losses in salmonid aquaculture worldwide but the influence of this bacterium on non-specific immunity in fish and the development of disease is uncertain. In our studies rainbow trout were diagnosed with gill infections caused by the bacterium after experiencing mortality in one of the fish farm’s tanks. For 6 weeks, we assessed the immunity parameters in affected fish. The activity of lysozyme and ceruloplasmin, as well as the level of total protein and immunoglobulins in fish serum were measured. Also, spleen phagocyte respiratory burst and potential killing activity, as well as head kidney lymphocyte proliferation stimulated by concanavalin A or lipopolysaccharide were determined. Almost all tested immunological parameters except for ceruloplasmin activity were reduced compared to the control group. F. psychrophilum seems to have mechanisms that allow it to evade the fish's immune system and suppress the basic non-specific humoral and cellular defense mechanisms of the infected fish.

Porcine parvovirus disease is a reproductive disorder caused by the porcine parvovirus (PPV) in sows and is characterised by miscarriage, stillbirth and mummification in pregnant sows. Porcine parvovirus disease poses a significant threat to pig herds and seriously hinders healthy and sustainable development of the pig farming industry. Currently, there is no effective treatment for porcine parvovirus disease except for prevention and control measures. Based on genotype differences, PPV can be classified into at least eight subtypes, PPV1-PPV8. Epigenetic mechanisms, particularly cytosine methylation of cytosine-phosphate-guanine (CpG) dinucleotides, are proven to have a significant impact on the life cycle of various viruses. Therefore, we selected the PPV genome as the research object and analysed the number, distribution and length of CpG islands in the genome of strains PPV1-PPV8. PPV1-6 had AT rich genomes (GC content ≤50%), whereas PPV7 had a GC content >50%. PPV1, PPV4, PPV5 and PPV6 contained fewer CpG islands (1-5), PPV7 contained moderate CpG islands (6-11) and PPV2 and PPV3 contained more CpG islands (12-16). This study provides a foundation for exploring novel antiviral treatment strategies from an epigenetic perspective.

For the current investigation, 5-7-year-old Kangayam bulls were used. The semen was collected twice in a week and two ejaculates in each time were collected. They were subsequently transported to the laboratory for processing of semen and maintained in a water bath at 34°C. On the day of semen collection, three groups of semen prepared with Tris-egg-yolk Glycerol Extender (TEYG) (group I), TEGY extenders with hyaluronan (group II), and TEGY extenders with metformin (group III) and stored in a water bath at 34°C. According to the group, the semen sample was first diluted in the ratio of 1:1 with the appropriate extender (TEYG, hyaluronan enriched or metformin enriched) and kept in laminar air flow at 22°C for seven minutes. Then each semen sample was extended using the appropriate semen extender in accordance with the dilution rate. After filling, sealing and printing, the final diluted semen sample was subjected to equilibration. As per the standard routine protocol freezing was done. Oocytes were collected from cyclical animals on days 1, 5, 9 after estrus by ultrasound guided transvaginal ovum pick-up method after ablation of day 0 pre-ovulatory follicle. After oocyte and sperm maturation, the co-incubation of oocyte and sperm was done and in vitro penetration rate was recorded. The overall in vitro penetration rate recorded in Kangayam cow was 46.66 per cent. Maximum in vitro penetration rate was observed in group II (52 per cent) followed by group III (46 per cent) and in group I (42 per cent). Based on the above findings, it was concluded that hyaluronan enriched semen may be used as pragmatic approach for cryopreservation of Kangayam bull semen in order to augment the in vitro penetration rate in Kangayam.

The aim of this study was to investigate the activity of thymoquinone (TQ), carvacrol (CAR) and thymol (TYM) against multi-drug resistant nontuberculous mycobacteria (MDR-NTM), alone and in combination with berberine (BER). Antimicrobial activity was first evaluated at concentrations from 8 to 512 µg/mL. Each of the compounds tested exhibited good activity against nontuberculous mycobacteria (NTM) isolated from fish, with MIC values of 32-128 µg/mL. In this study, we have shown for the first time the synergistic efficacy of BER with CAR, TYM or TQ against NTM strains. Thus, the combination of these compounds with BER seems to be a new approach for combating MDR-NTM strains.

Rhodococcus equi (R. equi) is a primary cause of pyogranulomatous pneumonia of foals between three weeks and five months of age. Early diagnosis of rhodococcal pneumonia has always been considered a preferable approach as it can lead to more successful treatment and better outcomes. In horse stud farms where the disease is common, neonatal foals are subjected to tests such as complete blood count and fibrinogen analysis at certain intervals. However, new biomarkers are needed in addition to blood count and fibrinogen measurement in this field for early diagnosis of diseases. Based on this need, in this study, the diagnostic importance of CL-11, SP-A, SP-D was investigated for the early diagnosis of pneumonia in foals naturally infected with R. equi. In a case-control design, fourteen 1-5 months-old foals with proven R. equi pyogranulomatous pneumonia and 10 healthy 1-5 months-old control foals were enrolled in this study. The median white blood cell count (WBC) and fibrinogen concentration in the case group were significantly higher than in the control group. The median CL-11, SP-A, and SP-D concentrations in the case group were also significantly higher than in the control group. However, there were overlaps in concentrations between groups for SP-A and SP-D. There was less overlap between the groups for the CL-11 concentration. The CL-11 assay was sufficiently accurate but over-diagnosed R. equi infection. The correlation plot between fibrinogen and CL-11 concentrations shows that this problem may be solved if CL-11 is used as the first biomarker and fibrinogen as the second check for those foals with CL-11 serum levels >0.8 and ≤1.5 ng/mL. As a result, in this study, it is recommended to use CL-11 together with fibrinogen to obtain more accurate results in diagnosing pyogranulomatous pneumonia caused by R. equi in foals on large horse stud farms.

Bisphenol A (BPA), an endocrine disrupting chemical, is an environmental toxicant widely used in the production of polycarbonate plastics, epoxy resins and paints. Ganoderma lucidum (GDL) is a plant with biological activities widely used in Chinese medicine. The present study aims to determine the effects of GDL against testicular dysfunction in rats exposed to BPA. For this purpose, a total of 24 Sprague Dawley rats, 6 rats in each group, were used in the study. Rats were administered 25 mg/kg/bw BPA and 300 mg/kg/bw GDL by oral gavage for 8 weeks. After the treatments, the rats were sacrificed and testicular tissues were removed. One of the testes was used for biochemical analyses and the other for histopathologic examinations. The caudal part of the epididymis was trimmed and semen was obtained. As a result, BPA increased MDA level in blood and testicular tissue, while it decreased CAT, GPx activity and GSH level. GDL treatment provided protection of the impaired oxidant balance (p<0.001). Furthermore, BPA caused decreased epididymal sperm motility and density, vesicular seminalis weight and blood testosterone levels, increased testicular and epididymal tissue weight (p<0.001). Histopathological examination revealed that BPA caused narrowing in testicular tubules and apoptosis, decreased Germinal cell thickness and androgen receptor number. It was determined that GDL administration preserved testicular histology. As a result, it was determined that BPA caused toxicity in the testicular tissue of rats, whereas GDL administration was ameliorative.

The effect of feeding spray dried porcine plasma (SDPP) to male turkeys during the first 4 wks of life was evaluated at 20 wks of age. A total of 648 male Hybrid Converter day-old turkeys were divided into two groups of 9 replicate pens of 36 birds each and fed a commercial-type program of 7 diet phases. In phase 1 birds were fed a corn-soybean meal Control diet for 4 wks, or a diet with 2% SDPP of similar nutrient density. Thereafter, both groups were fed common diets until the end of the trial at 20 wks of age. At 4 wks of age, birds were subjected to heat stress and crowding for a period of 24 hrs to simulate the stress induced commercially when moved from the brooder house into a grow-out building. Poults fed SDPP had greater body weight (BW) and body weight gain at 6 (P<0.05) and at 9 wks of age (P<0.10). At 20 wks of age, BW of turkeys fed SDPP vs Control was not different. However, the European Productivity Index (EPI) that incorporates final body weight, feed conversion ratio and livability, tended to be higher (P<0.10) for the SDPP group (EPI = 545 vs 529, respectively) because of the higher livability trend exhibited in SDPP group (96.91 vs 94.75%, respectively). Likewise, birds fed SDPP showed significantly lower frequency of foot pad lesions vs Controls (P<0.05) by the end of the study. In conclusion, SDPP fed early in life can reduce stress, improve performance and reduce incidence of foot pad lesions in turkeys.

In our experiment, we focused on continuous monitoring of the immunocontraceptive effect of Improvac® vaccine on the sexual activity of male goats, which was determined by measuring plasma testosterone levels, testicular biometric and ejaculate examination. The animals in the experimental group (n = 12) were administered two doses of 2 ml of Improvac® in a four-week interval; the animals in the control group (n = 5) received 2 ml of saline, respectively. Blood collection, semen collection and testicular measurements were performed at 14-day intervals. A total of 8 samples were collected from each animal. In 9 animals a significant decrease (p <0.05) in testosterone concentration was observed two weeks after the first dose. At the end of the experiment (16 weeks), eight goats reached the testosterone concentration below the detection limit, one goat had a concentration of 0.47 nmol/L. The testicular size was significantly (p <0.01) smaller four weeks after the first dose. At the end of the experiment, the testicular size was approximately three times smaller (p <0.001). Motility was 0% in two goats at the end of the experiment, 1 % in one animal and 10 % in one animal, respectively. The median sperm concentration was significantly lower (p <0.01) at the end of the experiment. A significant (p < 0.0001) shift in the percentage of morphological changes was recorded eight weeks after the first administration. At the end of the experiment, there were five animals with azoospermia, two with 100% morphologically altered sperm, one with 99% and one with 96% morphologically altered sperm, respectively. In the 3 male goats, a significant skin reaction occurred after the first application, which resulted in an inadequate response of the treatment. Our results show that Improvac has a significant effect on the sexual function and sperm production in 9 out of 12 bucks.

In homeostasis, which plays an important role in the proper functioning and maintenance of the internal functioning of the body, kidneys play a key role in being responsible for the proper homeostasis of glucose. Among glucose transporters sodium-dependent glucose co-transporters (SGLT’s) have a major role of the kidney‘s ability to reabsorb glucose. Despite the localization of these transporters has been extensively studied in mammals, there are still gaps in knowledge of the localization of SGLT‘s in birds of different age groups. The aim of the study was to immunolocalize in kidneys of hen chicken of different ages the sodium-dependent glucose co-transporters SGLT1 and SGLT2 comparing the localization between different age groups. The kidneys derived from 32 hen chickens (Gallus gallus domesticus) were devided equally into four age groups: 3, 7, 14, and 20 days old broilers, 8 indivuals in each group. In the study, the polyclonal primary antibodies Rabbit anti- SGLT1 and Rabbit anti-SGLT2 (Abcam, UK) were used together with the corresponding IHC kit (Abcam, UK). The results were visualized by taking photos with the AxioCam HRc camera (Germany) connected to the microscope Zeiss Axioplan-2 Imaging (Germany). The study revealed the similar immunolocalization of SGLT1 and SGLT2 in the apical part of cells of proximal renal tubules in hen chickens’ kidneys in all age groups. Strong staining of SGLT2 was noted also in the cytoplasm of epithelial cells of the proximal straight and convoluted tubules. Based on our study, the kidney tissue of newly hatched chicken is ready immediately after hatching for glucose reabsorption and transport, similarily to that of three-week-old chicks.

The material for drug resistance testing was 28 strains of Mycobacterium caprae isolated from tissue collected post mortem from a free-living Bieszczady Mountain European bison (Bison bonasus caucasicus) herd. All drug susceptibility tests were carried out on an automated Bactec mycobacterial growth indicator tube (MGIT) 960 system, using Bactec MGIT 960 streptomycin, isoniazid, rifampin and ethambutol (S.I.R.E.) and Bactec MGIT 960 PZA kits. The analyzed M. caprae strains demonstrated susceptibility to PZA and the complement of four basic anti-mycobacterial drugs: S.I.R.E. Considering that we are dealing with multidrug-resistant and extremely drug-resistant tuberculosis more and more often, and that no new drugs have been discovered or developed for over 60 years, the study of drug resistance in free-living animal strains of MTBC is of great importance for the deepening and broadening of our knowledge of TB.

This study analysed the influence of montelukast (MON), a cysteinyl leukotriene receptor antagonist and nifedipine, an L‐type voltage‐gated Ca2+ channel blocker, on the contractility of the porcine uterine smooth muscle. Myometrial strips were collected from the sexually immature (n=8), cyclic (12–14 days of the oestrous cycle; n=8) and pregnant (27-28 days of pregnancy; n=8) gilts and stimulated with a) MON or nifedipine at concentrations of 10-8-10-4 M and b) increasing concentrations of nifedipine after previous administration of MON at a concentration of 10-4 M. The changes in the tension, amplitude and frequency of contractions were determined with Hugo Sachs Elektronik equipment for measuring isometric contractions. Stimulation of the uterine strips with high concentrations of MON significantly reduced the amplitude of contractions in both the cyclic and pregnant group and the frequency of contractions in the pregnant group. In high concentrations, nifedipine significantly decreased the amplitude and frequency of contractions in all examined groups. Nifedipine administered after MON pre-treatment significantly decreased the tension in the pregnant group and the amplitude and frequency of contractions in all examined groups; this effect was more evident in comparison with nifedipine used alone. The obtained results indicate that the influence of MON and nifedipine on the contractile activity of the porcine uterus is dependent on the physiological status of the animal. Moreover, the blockage of the CysLT1 receptor enhances the relaxing effect of nifedipine.

The aim of this study here to evaluate the association between spontaneous echocardiographic contrast (SEC) and left atrial parameters such as size, volume, and function in cats with hypertrophic cardiomyopathy. Cats were assigned into groups; clinically healthy cats (n:8), HCM without SEC (n:12) and HCM with SEC (n:8) in the study. LAFS% and LAFAC had a statistical significance between groups. In conclusion, compared to healthy individuals, the presence of decreased trend in levels of LAFS% and LAFAC in HCMSEC cats could be a predictive marker for the thromboembolic risk assessment. Further studies enrolled the asymptomatic HCM cats in study groups need to be conducted to define a cut-off value of LAFS% and LAFAC before the SEC formation.

The outbreak and prevalence of tembusu virus (TMUV) endanger the breeding industry of waterfowls. However, little is known about the molecular mechanism underlying TMUV infection. It was reported that heat shock protein 70 (HSP70) was a positive regulator of the infection of TMUV. In order to study the interactions between HSP70 and host immune response to TMUV infection, TMUV-infected cells with or without HSP70 inhibitor were harvested and subjected to deep sequencing to identify genes differentially expressed. We found 43 differentially expressed genes (DEGs) in HSP70 inhibitor-treated and mock-treated TMUV-infected DF-1 cells. Of these DEGs, 39 genes were down-regulated significantly. Gene Ontology analysis suggested that the DEGs were mainly involved in biological process, cellular component and molecular function. Kyoto Encyclopedia of Genes and Genomes analysis showed that the DEGs mainly related to the activation of innate immune response, including RIG-I-like receptor, toll-like receptor and NF-κB signaling pathway. Also, 12 down-regulated immune-related DEGs were selected for confirmation by reverse transcription quantitative real-time PCR verification, all these genes showed consistent expression between the result of reverse transcription quantitative real-time PCR and transcriptomic sequencing. These results revealed the important role of HSP70 in facilitating the innate immune response induced by TMUV infection. This is first to access the role of HSP70 in host response to TMUV infection, which provides a basis for further study of the pathogenesis of TMUV and contributes to the elucidation of TMUV-host interactions.

Salmonella spp. is an important zoonotic and foodborne pathogen. It is spread worldwide and represents a public health risk. Pigs are a significant reservoir and are frequently subclinical carriers. The aim of this study was to detect the occurrence and antimicrobial resistance of Salmonella isolates, respectively the five most important for public health Salmonella serovars in fattening pigs in Bulgaria. The isolation of Salmonella spp. was carried out according to EN ISO 6579-1 for the detection of Salmonella bacteria in faeces and Salmonella serotyping following the Kauffmann-White scheme. All confirmed S. enterica isolates were analysed by the disk-diffusion method for susceptibility to 14 antimicrobials. Salmonella enterica was detected in 13 out of 32 tested farms in Bulgaria. The overall percentage of Salmonella positive pooled faecal samples was 6.8% (43 of 630 samples). The highest occurrence was present in fattening pigs aged between 121-180 days (16.3%; 20/123), followed by dry sows (6.5%; 4/62) and gilts (4.8%; 12/248). About 75% of the isolated strains belonged to three serotypes: Salmonella Infantis (41.9%), Salmonella Give (16.3%) and Salmonella Typhimurium monophasic (16.3%). All the tested isolates were resistant to Tilmicosin (100%), 88.4% to Ampicillin, followed by 69.8% to Tiamulin, 25.6% to Amoxicillin and Chlortetracycline. Multidrug resistance was recorded in 62.8 % of the tested strains. This study reports data regarding the circulation of five most important for public health Salmonella serovars (S. Enteritidis, S. Typhimurium, S. Typhimurium monophasic, S. Infantis and S. Derby) in farrow-to-finish pig farms in Bulgaria and represent 74.4% of the total Salmonella spp. isolates. This phenomenon has critical effects for the health on consumers and therefore represents a key “one health” issue.

Porcine alveolar macrophages（PAMs） can resist infection caused by Mycoplasma hyopneumoniae (Mhp) through phagocytosis. However, it is unknown what gene expression changes occur in PAM after Mhp stimulation. So differential gene expression (DGE) profiling technique was employed to analyze differentially expressed genes in PAMs infected with Mhp strain 232. eighty-six and 889 differentially expressed (DE) gene were identified in PAMs at 12 hours post-infection (hpi) and 24 hpi. By Gene Ontology (GO) analysis, DE genes were involved in 54 (12 hpi) and 128 (24 hpi) GO enrichment items. By Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis, DE genes were involved in 101 (12 hpi) and 250 (24 hpi) KEGG enrichment items. By Ingenuity Pathway Analysis (IPA) analysis, DE genes were connected, forming 25 internally interacting subnetworks. STRING analysis revealed 131 proteins encoded by DE genes involved in network interactions. Five novel genes were closely related to clinical symptoms and pathological changes of mycoplasma pneumonia in swine. It is the first report to investigate PAM transcriptional responses to Mhp infection by the DGE profiling technique.

Bovine endometritis has become a persistent issue in the global dairy business, resulting in huge economic losses. Due to their numerous positive benefits, Chinese herbal medicines (CHMs) have recently demonstrated remarkable pharmacological potential against endometritis. The objective of this study was to investigate the effects and elucidate the underlying mechanisms of the Yimucao formula (YMF) consisting of five herbs in lactation cows under endometritis conditions. Initially, the possible effects of YMF on cows with endometritis were assessed. Then, using network pharmacology, potential molecular processes by which the YMF prevents endometritis were suggested. The findings demonstrated a considerable improvement in endometritis-related clinical complaints following YMF treatment. Mechanically, 150 active compounds were identified from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP); of these, quercetin, kaempferol, beta-sitosterol, apigenin, isorhamnetin, and sitogluside were the most prevalent active substances. The NCBI gene, GeneCard, and OMIM databases had 110 genes linked to endometritis. The intersection of these targets with the 213 targets of the active ingredient produced 17 common targets, of which BCL2, IL-6, MMP9, HIF1α, TNF, IL-1β, and ICAM1 were the top 7 core targets. According to the Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment data, atherosclerosis, fluid shear stress, and the AGE-RAGE signaling pathway are the primary causes of YMF's anti-endometritis action. Finally, our results indicate that the YMF works on endometritis through various and multi-targeted signaling pathways, which aid in providing reference for clinical practice, based on network pharmacology and molecular docking.

Ultrasonographic renal length-to-aorta ratio is an ideal reflection of renal health in dogs. Earlier studies have highlighted the need for breed-specific reference intervals for this parameter. The aim of this study was to establish a breed-specific reference interval of kidney length to aorta ratio (KL:Ao) in Chippiparai dogs, a breed of Indian sighthounds. The KL:Ao ratio was recorded in 45 Chippiparai dogs, classified into 3 age groups. A narrow breed specific range of 7.07 to 7.74 was arrived at for adult Chippiparai dogs, and a strong relationship between age and body weight and KL:Ao was observed by regression analysis. These findings can be extrapolated and utilized by other sighthounds.

Flukes can cause severe and lethal diseases in various animals, including fish. Both adult and larval stages of flukes are found in fish. Haplorchiasis is an infection of fish gills by heterophyid trematodes such as Haplorchis taichui. To detect this parasite, the gills of 30 Tigris kingfish and Tigris barb collected from the Shapour River in Kazerun, Fars province were found to be parasitized with metacercarial cysts of a heterophyid trematode identified as H. taichui. Histopathological examination of the infected fish gills revealed cartilage proliferation, severe hyperplasia, fusion, S-forming, shortening and thickening, distortion, and displacement of affected secondary gill filaments leading to deformities of the filament structure, clubbing, telangiectasis, and hyperemia. Although the gill damage was evident and potentially life-threatening for the cyprinid fish, the examined fish showed no clinical signs. This .finding indicates that H. taichui is pathogenic; therefore, prevention of infection and treatment should be a priority.

Blastocystis is an intestinal protist commonly found in humans and many different animal species. It is probably the most common enteric parasite with an estimated one billion infections worldwide. The fecal materials for this study were collected from 100 cats and 200 dogs different age and sex in shelter in Van, Turkey. DNA extraction, PCR amplification and sequence analysis were performed on the fecal samples. As a result, a prevalence of 1% (1/100) in cats and 1.5% (3/200) in dogs was detected. The prevalence was higher in both cats and dogs, in age groups younger than one year and in females according to gender. Sequence analysis revealed Blastocystis sp. ST10 in cats and Blastocystis sp. ST1, ST10 and ST30 in dogs. The sequences obtained were deposited in Genbank. In conclusion, stray cats and dogs may be a source of infection for other cats and dogs, and the detection of zoonotic ST1 in dogs suggests that dogs may be a reservoir for human infection.

The present study aimed to detect the prevalence, virulence and antimicrobial resistance genes profile of E. coli isolated from diarrhoeic lambs. A total of 61 faecal samples were collected from diarrhoeic lambs. The presence of various virulence and antimicrobial resistance genes in E. coli isolates was determined by the use of PCR. In total, 46 E. coli isolates were recovered from 61 rectal swabs of diarrhoeic lambs. Out of these 46 isolates, PCR showed that seven isolates (15.22%) carried the stx1 or stx2 gene and were found positive for Shiga-toxin- producing E. coli (STEC). Four isolates (8.70%) were found to be Enteropathogenic E. coli (EPEC) and all these EPEC isolates were atypical EPEC pathotypes. STEC and intimin-positive isolates were recovered only from one isolate, hence, out of 46 isolates, only one isolate (2.17%) was confirmed as Enterohaemorrhagic E. coli. The lt and st genes were not detected in any of the E. coli isolates recovered from field samples. Therefore, all the isolates were confirmed as non-Enterotoxigenic E. coli. Further, Thirty-five isolates (76.09%) were found to be Entero-aggregative E. coli pathotypes. All the E. coli isolates were also tested for antimicrobial resistance against 15 different antibiotics. All the E. coli isolates were found to be resistant to penicillin-G, cephalothins, and azithromycin and the majority of isolates of E. coli were sensitive to chloramphenicol, ofloxacin, and sulfafurazole. Two antibiotic resistance genes i.e. tetA and blaTEM were detected in 10.87% (n=5/46) and 28.26% (n=13/46) of E. coli isolates respectively.