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.     

First Report on the Frequency and Molecular Detection of Neuropathogenic EHV-1 in Turkey

There has been an increase in outbreaks of neuropathogenic equine herpesvirus-1 (EHV-1) in the United States and Europe. However, the presence and frequency of neuropathogenic EHV-1 in Turkish horses are not known at present. This study aimed to investigate the frequency of EHV-1 and neuropathogenic strains of EHV-1 in the Marmara Region of Turkey. Samples were analyzed for the presence of EHV-1 and neuropathogenic EHV-1 by real-time PCR TaqMan probe assays. Overall detection rate of EHV-1 was 45.5% (51 of 112). The detection rates were 70.5% (24 of 34) in aborted fetuses, 53.3% (8 of 15) in neonatal deads, 66.6% (4 of 6) in foals, 40% (2 of 5) in dead mares, and 25% (13 of 52) in living mares. Overall detection rate of neuropathogenic EHV-1 was 7.8% (4 of 51), and the real-time PCR results were confirmed by sequencing. Neuropathogenic strains of EHV-1 were detected in the brain and lung of two mares with neurological disease but without a history of abortion, in the brain of a foal that died of respiratory disorder, and in the nasal swab from a mare with a history of abortion. On histopathology, nonpurulent meningoencephalitis, hemorrhages, and vasculitis were seen in the brain. In conclusion, results of this study indicated, for the first time, that the neuropathogenic EHV-1 is circulating in the Marmara Region of Turkey. The results of this study also show that the current risk for non-neuropathogenic strains is high, whereas risk for the neuropathogenic EHV-1-G₂₂₅₄ strain seems to be low. As outbreaks of EHV-1 continue in the Marmara region of Turkey, surveillance for neuropathogenic EHV-1 genotype should be maintained.     

Comparative Effects of Fish and Cow Gelatins and Locust Bean Gum on Chemical,Textural, and Sensory Properties of Yogurt

ABSTRACT

Yogurt is a traditional dairy product widely consumed all over the world. Syneresis, which is a result of lower water holding capacity, is a big concern and an important quality parameter. In this study, the efficiency of locust bean gum and different gelatins (fish skin, fish scale, and bovine skin) in preventing syneresis was investigated. Samples were stored at 4°C for 2 weeks, a typical shelf life of yogurt, and analyzed periodically for quality parameters. Syneresis, viscosity, and water holding capacity of the control and gum added samples were almost identical, while gelatin added samples were generally significantly improved (p < 0.05).     

Characterization of myofibrillar adenosine triphosphatase activity and liberation of actin from myofibrils upon heating chicken breast meat

Denaturation of actin and myosin in myofibrils induced by heating at 50°C was investigated to reveal the mechanism of irreversible liberation of actin from myofibrils on heating at lower temperatures than conventional cooking. Denaturation of these proteins was determined by Mg²⁺‐ATPase (adenosine triphosphatase) and Ca²⁺‐ATPase activities. When minced meat was heated for 20 min, actin was liberated accompanying denaturation of 80% of actin and 50% of myosin. Heating of the myofibrillar fraction (MFF) isolated from meat homogenate induced much slower denaturation of actin than myosin. When MFF was heated with sarcoplasmic fractions, denaturation of actin was facilitated, suggesting that sarcoplasmic fractions contain factors to facilitate actin denaturation. Inosine‐5′‐monophosphate, a component of sarcoplasmic fractions, was shown to have no effect on actin and myosin denaturation. These results suggest that heating meat at 50°C dissociates binding (‘Bond A’) between actin and myosin participating in ATPase activities, resulting in denaturation of both proteins under influence of sarcoplasmic components. Although denaturation of actin and myosin disrupted Bond A, actin was not liberated simultaneously, suggesting the presence of another bond (‘Bond B’, more heat‐stable than Bond A) between both proteins and necessity of disruption of Bond B for actin release from myofibrils.     

Molecular pathogenesis of bovine paratuberculosis and human inflammatory bowel diseases

Paratuberculosis (Ptb), caused by Mycobacterium avium subsp. paratuberculosis (Map), is a chronic enteritis that affects many ruminants and other wild animals worldwide. Ptb is a great concern in animal health and in etiology of human Crohn's disease (CD). In the present study, we detected Map-specific insertion sequence IS900 of DNA in tissue sections surgically removed from lesions of patients with CD (29 samples), ulcerative colitis (UC) (17 samples), and non-inflammatory bowel disease (IBD) (20 samples). We then compared the histopathological findings of 29 CD and 17 UC cases with those of 35 cases of bovine Ptb, since few comparative pathological studies of human IBD and Ptb have been conducted. The QPCR examination indicated positive results in 13.37% of CD cases, 3.57% of UC cases, and 10% of non-IBD cases. Human CD tissues typically exhibited destructive full thickness enteritis with severe lympho-plasma infiltration and scattered additional granulomas; UC lesions exhibited much less inflammation than CD lesions. Non-IBD control samples did not exhibit pathological changes. Human CD and UC lesions were very different from Ptb lesions that are characterized by predominant granuloma formation. Immunohistochemistry for Map antigen and acid-fast staining were negative in all human IBD cases but were always positive in Ptb cases. Our present comparative study strongly suggests that we reconsider the previous hypothesis that "Map infection" causes CD, even though human intestines were considered to have been exposed to the Map antigen containing the DNA.     

Consensus-based reporting standards for diagnostic test accuracy studies for paratuberculosis in ruminants

The Standards for Reporting of Diagnostic Accuracy (STARD) statement (www.stard-statement.org) was developed to encourage complete and transparent reporting of key elements of test accuracy studies in human medicine. The statement was motivated by widespread evidence of bias in test accuracy studies and the finding that incomplete or absent reporting of items in the STARD checklist was associated with overly optimistic estimates of test performance characteristics. Although STARD principles apply broadly, specific guidelines do not exist to account for unique considerations in livestock studies such as herd tests, potential use of experimental challenge studies, a more diverse group of testing purposes and sampling designs, and the widespread lack of an ante-mortem reference standard with high sensitivity and specificity. The objective of the present study was to develop a modified version of STARD relevant to paratuberculosis (Johne's disease) in ruminants. Examples and elaborations for each of the 25 items were developed by a panel of experts using a consensus-based approach to explain the items and underlying concepts. The new guidelines, termed STRADAS-paraTB (Standards for Reporting of Animal Diagnostic Accuracy Studies for paratuberculosis), should facilitate improved quality of reporting of the design, conduct and results of paratuberculosis test accuracy studies which were identified as "poor" in a review published in 2008 in Veterinary Microbiology.     

Geographical variation of foxtail millet, Setaria italica (L.) P. Beauv. based on rDNA PCR–RFLP

The rDNA PCR–RFLP of foxtail millet (Setaria italica) germ-plasm collected throughout Eurasia and from a part of Africa was investigated with five restriction enzymes according to our previous study. Foxtail millet germ-plasms were classified by length of the rDNA IGS and RFLP; clear geographical differentiation was observed between East Asia, the Nansei Islands of Japan-Taiwan-the Philippines area, South Asia and Afghanistan-Pakistan. We also found evidence of migration of foxtail millet landraces between the areas. We calculated diversity index (D) for each region and found that center of diversity of this millet is East Asia such as China, Korea and Japan.     

Effect of varying dietary energy level on feed intake, feed conversion, whole-body composition and growth of Malawian tilapia, Oreochromis shiranus– Boulenger

Four isonitrogenous [30% crude protein (CP)] diets containing different gross energy levels (13.39, 16.74, 20.50 and 23.85 kJ g⁻¹) were evaluated to determine the optimum energy for the Malawian tilapia Oreochromis shiranus. Each tank (120 L) was stocked with 18 juvenile tilapia (average weight 7.32±0.25 g) and they were fed the experimental diets for 10 weeks. The final average weight of the fish was approximately twofold higher (range: 12.64–16.77 g) than the initial weight. The dietary energy significantly (P<0.05) influenced growth. The average weight of fish fed dietary energy level 20.50 kJ g⁻¹ was significantly higher (P<0.05) than the weight of the fish fed any of the other experimental diets. There was no significant difference in growth of fish fed 13.39 and 16.74 kJ g⁻¹ energy levels, but 23.85 kJ g⁻¹ produced the lowest growth rates. There were no significant differences (P>0.05) between feed intake across the treatments. Feed conversion ratio (range: 2.2–3.0) and protein efficiency ratio (range: 1.10–1.50) among the dietary treatment groups were in agreement with trends for weight gain. Dietary energy level significantly (P<0.05) influenced the body composition of O. shiranus. Whole‐body moisture (range: 64.27–67.15%) and ash (range: 13.21–14.73%) decreased in all treatments. Whole‐body protein (range: 63.57–66.16%) increased only in groups fed on the diet containing 20.50 kJ g⁻¹. Whole‐body fat (range: 13.58–17.27%) and gross energy (range: 28.411–33.210 kJ g⁻¹) increased significantly (P<0.05). Fish survival was 100% in all treatments. The results demonstrated that to maximize growth at a temperature of 23°C, O. shiranus should be fed diets containing 20.50 kJ g⁻¹ gross energy.