Veterinary Research Forum

Abstract Contagious caprine pleuropneumonia (CCPP) in goats is defined as a highly contagious and rapidly spreading mycoplasmal disease that is now among the leading causes of major economic losses on many continents (Asia, Africa and the Middle East). In this study, we aimed to evaluate immunohistochemically mast cells (MCs) profile and local interleukin (IL)-17 and IL-1β protein expressions in naturally infected CCPP according to the course of the inflammation (peracute-acute, subacute-chronic). The material of the study consisted of 40 naturally infected CCPP and 6 healthy control goat lung tissues. Appropriate samples were taken from the necropsied goats and subjected to histopathological and immunohistochemical examination. In the histopathological examination of the samples, it was determined that 29 samples had a peracute-acute course and 11 had a subacute-chronic course. In immuno-histochemical examination, MC profile and local IL-17 and IL-1β protein expressions were evaluated in the peracute-acute and subacute-chronic course. Immunohistochemically, significant increases in MC number, local IL-17 and IL-1β scores were detected in the peracute-acute course compared to the control group. There were significant decreases in the relevant scores in the subacute-chronic course compared to the peracute-acute course. Current findings indicated that MC, IL-17, and IL-1β expressions played important roles in the pathogenesis of infection in naturally infected CCPP, especially in the peracute-acute course. Additionally, MC profile was evaluated for the first time in naturally infected CCPP.

Abstract An increase in morbidity during times of relatively high functional load on the body such as during pregnancy, confirms the role of metabolic overload in the occurrence of metabolic failures. It is better to take preventive measures such as adjusting metabolic regulation mechanisms in light of the ideal dietary composition. However, this direction is constrained by the lack of information about neurohumoral regulation. The goal of the present study was to learn more about the dynamics of changes in insulin and glucose levels in pregnant cow’s blood. Research on the levels of glucose and insulin in lactating cows demonstrated that ruminants had distinct characteristics in the metabolism of carbohydrates, most notably a lessened reliance of blood glucose on insulin levels. A rise in blood glucose and insulin levels was observed as the gestational stage increased during the third trimester of pregnancy. The intensity of this insulin tolerance was contingent upon the level of productivity and glucose levels during the preceding stages of gestation.

Abstract This study was important to improve proper biosecurity measures and controlling the spread of Aeromonas to prevent future outbreaks. This research sought to determine whether virulent Aeromans species were present in morbid rainbow trout, their resistance and their genetic relatedness. A total number of 542 tissue lesion specimens were collected from gill, liver, heart and kidneys in morbid domesticated fish in Duhok province, Iraq. The gyrB DNA sequence analysis was used to determine the species classification. Drug susceptibility testing was conducted for all isolated strains using disc diffusion technique. The genotyping analysis was carried out using enterobacterial repetitive intergenic consensus-polymerase chain reaction. Thirty-four isolates were found and they were classified into three species (Aeromonas veronii, Aeromonas sorbia, and Aeromonas allosaccharophila), where A. veronii stand as one of the most prevalent species. The most frequently affected organ by Aeromonas was the gills among four different organs. The detection frequencies of the virulence genes aerolysin, outer membrane protein, glycerophospholipid-cholesterol acyltransferase, elastase, flagella, serine protease, cytotonic heat-labile, and hemolysin were 100%, 100%, 79.41%, 64.70%, 76.47%, 67.64%, 70.58%, and 41.17, respectively. None of the strains possessed all of the virulence markers. All isolates were completely resistant to ceftazidime, amoxicillin and doxycycline. All isolates were found to be multi-drug-resistant. Regardless of the nearest geographic source area of samples and the same Aeromonas species, there was a high genetic diversity. The results of this study could help farmers and researchers make informed decisions about measures of biosecurity and proper therapeutic drugs to apply to prevent current outbreaks and prevent them from recurring again.

Abstract The ability of lactic acid bacteria (LAB) to produce biogenic amines (BAs) from amino acids using amino acid decarboxylase enzymes is an important food safety criterion due to their use as starter or bio-protective cultures. In this study, various LAB isolates, including Lactococcus (three isolates), Lactobacillus (five isolates), Enterococcus (13 isolates), and Leuconostoc (seven isolates), were isolated from ready-to-eat fish products like sauced, marinated, smoked anchovies, and smoked mackerel. These isolates were then tested for their ability to produce BAs (histamine, putrescine, cadaverine, spermidine, tryptamine, 2-phenylethylamine, spermine, serotonin, tyramine, 3-methylamine, and dopamine) and ammonia in histidine decarboxylase broth. The post-biotic solutions (cell-free supernatant) of Lb. paracasei NZ.Lbp.111, Lb. brevis NZ.Lbb.085, and E. casseliflavus NZ.Ec.074 exhibited the highest ammonia levels. Histamine accumulation was generally low in post-biotic solutions from four LAB isolates, ranging from 2.00 to 7.78 ppm. However, the post-biotic solution of Leu. mesenteroides NZLeu 009 displayed a significantly higher histamine concentration (83.23 ppm). Lactobacillus post-biotic solutions had the highest concentrations of ammonia (1220.28 ppm), 2-phenylethylamine (82.96 ppm), serotonin (278.70 ppm), tyramine (267.48 ppm), and cadaverine (19.72 ppm). Also, the average BAs concentration for Lactobacillus post-biotic solutions was the highest at 31.98 ppm. The results revealed the analysis of BAs concentrations in different LAB isolates from fish products, providing interesting insights into their metabolic capabilities and potential implications for food safety and quality.

Abstract Cryopreservation and re-transplantation of ovarian tissue is a relatively new technique to preserve fertility. This study evaluated the preservation of ovarian follicles after the vitrification-thawing procedure by implanting ovarian fragments into the gluteus muscle of rabbit. Bilateral ovariectomy was performed on each rabbit. The cortices were separated from the medullae of ovaries and divided into four fragments. One fresh cortical fragment was used as a control, fixed in 10.00% formalin. Three fragments underwent vitrification-thawing for two weeks. Two of these vitrified-thawed fragments were auto-transplanted into separate locations within the opened gluteus muscle, while the final fragment was fixed in 10.00% formalin. Eight weeks after re-implantation, biopsies were collected from the ovarian fragments and fixed in 10.00% formalin. The numbers and areas (µm²) of morphologically normal follicles were evaluated on sections stained with Hematoxylin and Eosin through light microscopy. The follicular morphology in the ovarian fragments immediately after vitrification-thawing remained similar to that of the fresh ovary. The number of morphologically intact follicles was significantly lower in the ovarian fragments eight weeks after re-implantation than fresh ovary and ovarian tissue fragments after vitrification-thawing procedure. However, follicular development to the antral stage was observed in all samples eight weeks after re-implantation. There was no statistical difference in the areas (µm²) of primordial, primary, pre-antral, and antral follicles in the ovarian fragments before cryopreservation, immediately after vitrification-thawing, and following re-implantation procedures. Auto-transplantation of ovarian tissue into the gluteus muscle of rabbits could be a viable approach for preserving ovarian follicles after vitrification-thawing procedure.

Abstract Aflatoxins are toxic chemicals produced by Aspergillus fungi. Reports exist on the relationship of aflatoxin exposure via contaminated food and feed to hepatotoxicity and liver cancer. Aflatoxin B1 (AFB1) and Aflatoxin G1 (AFG1) are two dangerous types of aflatoxins for human health. Bovine α-lactalbumin (ALA) is the second major whey protein in milk which bear diverse biological functions. In this study, the interaction of AFB1 and AFG1 with the ALA protein was studied using fluorescence spectroscopy, molecular docking and molecular dynamic (MD) simulation. The spectroscopy experiments showed that the interaction with AFB1 and AFG1significantly quenched the intrinsic fluorescence emission of ALA via a static quenching mechanism. The free energy of binding and binding constant (Ka) obtained from the intrinsic fluorescence results were –5.32 kcal per mol and 0.80 × 10⁴ L mol^-1 for AFB1 and -5.64 kcal per mol and 1.35 × 10⁴ l mol^-1 for AFG1, respectively. Molecular docking studies were conducted before and after the MD simulation to estimate the binding sites, Ka s and binding mode. Results from the molecular docking showed that AFB1 and AFG1 bound to ALA via hydrophobic interaction and hydrogen bond. After MD simulation, the precision of the Ka obtained from the docking results was improved and it was more similar to the experimental results of fluorescence spectroscopy. Other simulation results were aligned well with the molecular docking and fluorescence spectroscopy results. Accordingly, AFB1and AFG1 could form complex with ALA, however, AFG1 showed higher affinity for binding to ALA and more compact complex structure.

Abstract Hydropericardium syndrome (HPS) has caused significant financial losses to the Iranian poultry industry in the past few years. Thirty-two broiler chickens with gross lesions of HPS were inspected histologically and immunohistochemically. Sampling was performed in Sabzevar, Iran. The dead and sick birds from random farms were subjected to necropsy examinations. Only four broiler chickens had no hydropericardium and the other gross findings were similar for birds. Basophilic and eosinophilic intranuclear inclusions, hemorrhages and necrosis in different organs were the primary characteristic histologic lesions. Lymphoid depletion, goblet cell hyperplasia and necrotizing enteritis were some of the findings reported in previous research. Low macrophage infiltration rate and brain lesions were other discoveries in Hematoxylin and Eosin (H&E) examination. Feulgen reaction and Cluster of Differentiation 68 (CD68) immunohistochemical staining were used for a comprehensive investigation and these techniques revealed improved histopathologic details. Feulgen staining confirmed brain lesions and some other changes in different organs. Eventually, the CD68 method revealed low macrophage presence in most organs. This study suggested that HPS might cause brain damage and the susceptibility of the Arian breed to the adenovirus needs further investigation.

Abstract Leptospirosis is a worldwide zoonotic disease caused by pathogenic Leptospira spp., often occurring in tropical and subtropical regions. Focusing on development of rapid diagnostic methods to facilitate early diagnosis and a universal vaccine are the main critical issues to overcome the burden of leptospirosis. Here, we have studied the immunogenic potential of prepared recombinant Loa22 protein (rLoa22) of local pathogenic Leptospira species in mice and its ability to induce humoral and cellular immunity, being further evaluated by analyzing the immunoglobulin G (IgG) subclasses and cytokines produced through immunization. Based on the results, mice immunized with rLoa22/adjuvant and a trivalent vaccine, induced high titers of total IgG. All immunized groups increased IgG1 almost on the same level; but, IgG2a level was significantly higher in the vaccine and rLoa22/adjuvant groups than rLoa22 alone group. Animals immunized with the vaccine produced more interleukin 4 than rLoa22/ adjuvant group. The results of evaluating interferon gamma level showed that the rLoa22/adjuvant and vaccine groups had a significant increase compared to the rLoa22 alone group. The results also demonstrated that the rLoa22 protein, in indirect enzyme-linked immunosorbent assay, was able to detect the anti-Leptospira antibodies in mice serum that can be used as a marker in assessing the seroprevalence of leptospirosis and/or in combination with other leptospiral antigens in development of an effective vaccine against leptospirosis.