Aydın Gürel

Pathologic Alterations Detected in Acute Pancreatitis Induced by Sodium Taurocholate in Rats and Therapeutic Effects of Curcumin, Ciprofloxacin and Metronidazole Combination

Background and Aims: Secondary bacterial infections and free radical injury have been known to play an important role in the pathogenesis and clinical outcome of acute pancreatitis. Despite the therapy models developed in recent years, the mortality rate is still reported to be higher than expected. The objective of this study therefore was to investigate the effectiveness of ciprofloxacin and metronidazole combination and curcumin together in the treatment of acute pancreatitis. Methods: Acute pancreatitis was induced in rats by sodium taurocholate (n = 60). Starting 6 h after the induction of acute pancreatitis, groups I and II were injected 200 mg/kg ciprofloxacin and 500 mg/kg metronidazole intraperitoneally every 12 h for 6 days. Groups II and III received 100 mg/kg curcumin since day 20 prior to the initiation of acute pancreatitis. On day 6, animals of all groups were killed. Blood and tissue samples were taken for biochemical, pathologic and bacteriologic examination. Results: No statistical difference in the treatment groups versus the non-treatment group has been detected in the pancreatic tissue on the basis of histopathological scoring results. Prevalences of bacterial translocation were significantly lower in the treatment groups (groups I–III) than in the non-treatment group (group IV) (p < 0.001, p < 0.001, p < 0.05, respectively). Serum amylase, lipase, malon dialdehyde and nitric oxide (except for nitric oxide level in group I), levels of groups I, II and III were significantly lower than those of group IV (p < 0.05). Conclusions: The administration of ciprofloxacin and metronidazole in combination and curcumin in acute pancreatitis failed to provide a preventive effect on the occurrence of tissue injury, whereas free radical injury and prevalence of bacterial translocation were reduced significantly.

Abattoir study of maedi-visna virus infection in Turkey

MAEDI-VISNA virus is a lentivirus and a member of the Retroviridae family, which also contains other lentiviruses which cause immunodeficiences in domesticated animals and primates (Carey and Dalziel 1993, Pepin and others 1998). Maedi-visna virus infection was first reported as a cause of progressive pneumonia and progressive paralysis in sheep in Iceland in 1939. It now has a worldwide distribution, causing pneumonia, nervous system disorders, mastitis and arthritis (Cutlip and Laird 1976, Crawford and others 1980, Van der Molen and others 1985, Watt and others 1994, Dawson and Clarkson 1995, Pekelder and Clarkson 1995). Maedi-visna was first reported in Turkey in 1987 (Girgin and others 1987). The disease is chronic and most of the cases are subclinical; the disease is, therefore, usually seen in older animals. Affected animals also often have pulmonary adenomatosis and/or other infections caused by, for example, Pasteurella and Echinococcus (Pritchard and Done 1990, Watt and others 1992, Giangaspero and others 1993, Gonzales and others 1993). In maedi-visna infection, the lungs and associated lymph nodes are enlarged, and the lungs are congested and have

Lesions in the thymus and bone marrow in chicks with experimentally induced chicken infectious anemia disease.

One-day-old SPF chicks were inoculated with the Cux-l strain of chicken infectious anemia virus (CIAV), and the clinical development of disease and its macroscopic and microscopic alterations in the thymus and bone marrow, were observed. Tissue sections of thymus and bone marrow were stained using the streptavidin-biotin peroxidase method and examined under light microscope for evaluation of antigenic intensities in tissues. Those findings were then compared with blood parameters and ELISA results obtained through collected sera during sacrifice procedures. We sought to determine: the localization of viral antigens in thymus and bone marrow tissues after inoculation, the correlation between antigen intensities and hematologic, serologic and histopathologic findings, definitive diagnostic criteria using histopathologic and immunoperoxidase methods, and the reliability of these methods in the diagnosis of CIAV infection. For this purpose, 83, one-day-old SPF chicks were used. The birds were divided into experimental (n = 52) and control (n = 26) groups. A virus dose of TCID50 of 100,000/ml was administered intramuscularly to every bird in the experimental group. Based on the results of this study, we have suggested that clinical examination, along with macroscopic and microscopic evaluation of the thymus and bone marrow, maybe undertaken starting from day 7 post-inoculation (PI). ELISA, might be of value, as it might give consistent results starting from day 14 PI. However, the most reliable results were obtained through examination of thymus and bone marrow sections from infected birds stained by immunoperoxidase technique, as early as day 4 PI.

Efficacy of doxycycline release collagen membrane on surgically created and contaminated defects in rat tibiae: A histopathological and microbiological study

BACKGROUND The effects of systemic antibiotics on controlling infective pathogens after guided bone regeneration(GBR) procedures especially in membrane exposures are limited. However, local administrations of antibiotics are rare in GBR techniques. AIM The aim of this study was to investigate the osteogenesis potential and the antibacterial effect of a doxycycline releasing collagen membrane in surgically created and contaminated defects in rat tibiae. MATERIAL AND METHODS Defects were created in 20 rats that were randomly divided in to two groups: control group (defect contaminated by Porphyromonas gingivalis, filled with bone graft and covered by collagen membrane); test group (defect contaminated by P. gingivalis filled with bone graft and covered by collagen membrane containing 1mg/cm(2) doxycycline. Animals were sacrificed post surgically on the 14th day for microbiologic evaluation and on the 28th day for histopathological evaluation. RESULTS The degree of osteogenesis in the test group was seen to be significantly higher than control group (p: 0.011; p<0.05). Furthermore in test group, no bacterial growth was observed. The bacteria counts were determined between 1×104 and 268×104CFU/g with a median of 1.32×104 for control group. CONCLUSIONS Within the limitations of this study, the results of the present study suggests that the use of a doxycycline releasing membrane has a positive effect on contaminated GBR procedures for limiting P. gingivalis infections leading to bone formation following GBR procedures in a rat model.

Molecular and pathological investigations of EHV-1 and EHV-4 infections in horses in Turkey

The aim of the present study was to investigate abortion storms that occurred in the Marmara region of Turkey in 2008-2009 using a real-time PCR. Two aborted foetuses were necropsied and histo-pathological findings reported herein. Ten lungs, 3 brains and one nasal swab from 10 aborted foetuses, 6 nasal swabs and 3 vaginal swabs from aborting mares were included in this study. EHV-1 was isolated from the lung, liver and brain of 1 aborted foetus. EHV-1 DNA was detected in the lungs, livers and spleens of 2 necropsied foetuses and in 3 lungs from 10 foetuses submitted for diagnosis. A brain from one of the aborted foetuses was also positive for EHV-1 DNA. EHV-4 DNA was detected only in a nasal swab of one of the tested foetuses. Neither EHV-1 nor EHV-4 DNA was detected in the swabs of aborting mares. Sequence analysis of the glycoprotein B of the strains was performed and a phylogenetic tree was generated. The results indicated that 4 of the 5 Turkish EHV-1 strains (TR02, TR03, TR04 and TR05) clustered together; the fifth strain (TR01) was slightly removed from the group and clustered with other EHV-1 from various origins. Single nucleotide polyporphism (SNP in ORF30) associated with neuropathogenesis was not detected in any of the strains. At necropsy, sub-milier focal necrosis in the liver and spleen was observed. Microscopically, focal coagulation necrosis and marked eosinophilic intranuclear and intracytoplasmic inclusion bodies in the hepatocytes localised around the necrotic areas in the liver. Severe coagulation necrosis in white pulp of the spleen was also observed.

Frequency, clinicopathological features and phylogenetic analysis of feline morbillivirus in cats in Istanbul, Turkey:

Objectives The aim of the study was to investigate feline morbillivirus (FmoPV) frequency, phylogeny and associated pathology in cats in Istanbul, Turkey. Methods Samples from sick (n = 96) and dead (n = 15) cats were analysed using reverse transcription PCR. Blood and urine analyses and histopathology were also performed. Results FmoPV RNA was detected in six cats (5.4%), including three sick (in the urine) and three dead cats (tissues). A significantly greater proportion of FmoPV RNA-positive cats had street access compared with non-infected cats. Blood samples from the morbillivirus-positive cats were negative for morbillivirus RNA. Tubular parenchymal cells, lymphoid and plasma cells in kidney and hepatocytes, lymphoid and plasma cells in liver from dead cats were also positive by immunohistochemistry for the viral N protein. Two FmoPV-positive cats were also positive for feline coronavirus RNA and one cat for feline immunodeficiency virus RNA and feline leukaemia virus proviral DNA. Phylogenetic analysis of the six FmoPV-positive cats showed that the strains were grouped into cluster D and had high similarity (98.5–100%) with strains from Japan and Germany. In the three FmoPV RNA-positive sick cats, respiratory, urinary and digestive system signs were observed as well as weight loss, fever and depression in some cats. Similar clinical signs were also seen in the morbillivirus RNA-negative sick cats. FmoPV RNA-positive cats had lower median red blood cell count, haemoglobin, albumin, albumin/globulin and urobilinogen and higher alanine transaminase, alkaline phosphatase and bilirubin compared with non-infected cats. Significant histopathology of FmoPV RNA-positive dead cats included tubulointerstitial nephritis characterised by severe granular and vacuolar degeneration of the epithelial cells of the cortical and medullary tubules as well as mononuclear cell infiltrates. Widespread lymphoid cell infiltrates were detected in the renal cortex and medullary regions of the kidneys. Cellular infiltration, cholangiohepatitis and focal necrosis in the liver were also found. Although virus-infected cells were found in the kidney and liver of FmoRV RNA-positive cats, tubulointerstitial nephritis, cholangiohepatitis and focal necrosis seen in FmoRV RNA-positive cats were similar to those observed in FmoRV RNA-negative cats. Conclusions and relevance This is the first study to show the presence of FmoPV infection in cats in Turkey. Sick cats, particularly those with kidney disease, should be tested for this virus. The genotypes found in this study were similar to previously reported strains, indicating that circulating morbilliviruses in Turkey are conserved.

Phylogeny and S1 Gene Variation of Infectious Bronchitis Virus Detected in Broilers and Layers in Turkey

SUMMARY The avian coronavirus infectious bronchitis virus (AvCoV-IBV) is recognized as an important global pathogen because new variants are a continuous threat to the poultry industry worldwide. This study investigates the genetic origin and diversity of AvCoV-IBV by analysis of the S1 sequence derived from 49 broiler flocks and 14 layer flocks in different regions of Turkey. AvCoV-IBV RNA was detected in 41 (83.6%) broiler flocks and nine (64.2%) of the layer flocks by TaqMan real-time RT-PCR. In addition, AvCoV-IBV RNA was detected in the tracheas 27/30 (90%), lungs 31/49 (62.2%), caecal tonsils 7/22 (31.8%), and kidneys 4/49 (8.1%) of broiler flocks examined. Pathologic lesions, hemorrhages, and mononuclear infiltrations were predominantly observed in tracheas and to a lesser extent in the lungs and a few in kidneys. A phylogenetic tree based on partial S1 sequences of the detected AvCoV-IBVs (including isolates) revealed that 1) viruses detected in five broiler flocks were similar to the IBV vaccines Ma5, H120, M41; 2) viruses detected in 24 broiler flocks were similar to those previously reported from Turkey and to Israel variant-2 strains; 3) viruses detected in seven layer flocks were different from those found in any of the broiler flocks but similar to viruses previously reported from Iran, India, and China (similar to Israel variant-1 and 4/91 serotypes); and 4) that the AVCoV-IBV, Israeli variant-2 strain, found to be circulating in Turkey appears to be undergoing molecular evolution. In conclusion, genetically different AvCoV-IBV strains, including vaccine-like strains, based on their partial S1 sequence, are circulating in broiler and layer chicken flocks in Turkey and the Israeli variant-2 strain is undergoing evolution.

Aggravating effect of atorvastatin on indomethacin-induced gastric injury: Focus on PGE2, TNF-α, neutrophils and iNOS.

Statins are suggested to possess healing properties due to their antioxidant and antiinflammatory effects in animal ulcer models. In contrary, a clinical report indicated the formation of gastric ulcer by the use of atorvastatin. In this study, we aimed to investigate the effects of atorvastatin (0.5, 5 and 50mg/kg, p.o.) after single (acute) and multiple (subchronic, 5 days) applications on indomethacin-induced gastric ulcer in rats. In both acute and subchronic models high dose atorvastatin (50mg/kg), unlike to lower doses (0,5 and 5mg/kg), significantly aggravated ulcer lesions induced by indomethacin (30 mg/kg) although, a direct ulcerogenic influence was lacking. Proulcerogenic effect of atorvastatin are likely to be associated with decreased mucosal defense mechanisms (GSH and PGE2), and increased neutrophil infiltration and proinflammatory factors (TNF-a and iNOS) possibly via independently from mevalonate pathway. Thus, atorvastatin therapy should be monitorized in patients for an increased risk of gastric ulcer particularly when used concomitantly with NSAIDs.

Production of Recombinant N Protein of Infectious Bronchitis Virus Using the Baculovirus Expression System and Its Assessment as a Diagnostic Antigen

The avian coronavirus-infectious bronchitis virus (AvCoV-IBV) is recognized as an important avian pathogen, and new viral variants are a continuous threat to the poultry industry worldwide. Sensitive diagnostics and efficacious vaccines are necessary to combat IBV infections in chickens. The aim of this study was to produce recombinant N protein of IBV in the baculovirus system to use in ELISA diagnostic tests in order to enable the assessment of the sero-prevalence and risk of IBV infections in chickens in Turkey. For this, the gene encoding the N protein of the Beaudette strain of IBV was expressed using a recombinant baculovirus expression system. The recombinant N protein was purified using Ni-NTA affinity chromatography. An estimated 50-kDa recombinant protein corresponding to the expected molecular weight of IBV N including the 6xHis tag was detected using an anti-His monoclonal antibody. Specific immunoreactivity of the recombinant protein was confirmed by Western blot using antiserum obtained from vaccinated and naturally infected chicken from Turkey as well as using a monoclonal antibody raised against the N protein of the IBV Massachusetts strain. The results obtained with the in-house ELISA had high agreement with a commercial ELISA. Immunoreactivity analysis using antisera in Western blotting and the in-house ELISA suggests that the recombinant IBV N protein could be broadly cross-reactive with antisera produced against different IBV strains. We conclude that the recombinant baculovirus expressed IBV N protein could serve as a useful diagnostic antigen for detection of IBV infections in chickens by ELISA.

Clinical, Virological and Pathological Investigations on Horses with Neurological Disorders in Turkey

ABSTRACT The aim of this study was to investigate the clinical, pathologic, and viral etiology of horses with neurologic disorders. Twelve English Thoroughbred horses with neurologic disorders were investigated for the presence of neuropathogenic equine herpesvirus‐1 (EHV‐1) and Borna disease virus (BDV) by real‐time polymerase chain reaction (PCR) and histopathological methods. Neuropathogenic EHV‐1 was detected in the brain of two horses by real‐time PCR. Borna disease virus p24 and p40 gene sequences were detected by real‐time RT‐PCR in the brain of a 3‐year‐old horse and in the blood of a 1‐year‐old horse. High fever, ataxia, depression, lack of coordination, and gait abnormalities were present in these horses, which died within a few days of developing neurologic signs. The BDV p24 and p40 real‐time PCR products were sequenced and shown to be identical to previously reported BDV sequences. In the brain of the BDV‐positive horse, hyperemia was pronounced in the parenchyma and the meninges. In addition, nonpurulent poliencephalomyelitis characterized by perivascular mononuclear cell infiltration was seen. Mononuclear and polymorphonuclear cell infiltration was also seen in the liver with necrotic perihepatitis besides subacute splenitis and glomerulonephritis and severe hyperemia in the kidney and edema and emphysema in the lung. In conclusion, neuropathogenic EHV‐1 and BDV were detected in horses from Turkey using molecular detection methods. Since BDV genetic signatures were detected in Turkey for the first time, future epidemiologic studies need to be performed to investigate the range of host animals, spread, frequency, and molecular diversity of BDV; this will allow to determine the risk of this pathogen for Turkish veterinary and public health. HighlightsThis study describes the first cases of Borna disease in horses and first molecular detection of Borna disease virus in Turkey.