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Bovine rotavirus (BRV) which is the leading viral agent of neonatal calf diarrhea. In recent years, the use of shotgun metagenomics has emerged as a powerful tool for the detection and characterization of viruses. In the present study, BRV was isolated on MA-104 cells from feces of a neonatal calf with diarrhea and shotgun metagenomics was performed in order to obtain whole genome of BRV isolate. From nine sequenced segments, five segments encoding VP1, VP6, NSP3, NSP4, and NSP5/6 proteins was sequenced in full-length and four segments encoding the VP2, VP3, VP7, and NSP2 proteins was sequenced partially and are deposited in GenBank. Sequences of 9 out of 11 segments of BRV can be gained from metagenomics and the BRV isolate which is named as “RVA/Cow-tc/TUR/KIRSEHIR/2019/G10P[X]” is genotyped as G10-P[X]-I2-R2-C2-M2-AX-N2-T6-E2-H3. The VP1 and VP7 nucleotide and amino acid sequences of the present virus are conserved among Turkish BRV strains. Moreover, the VP2 gene/protein was genetically close to a number of RVA strains of cat and dog. Interestingly, for the NSP4 gene/protein, many equine rotaviruses from Japan and feline rotaviruses were found to be similar to KIRSEHIR strain. Although the highest nucleotide identity for NSP5/6 gene was with buffalo rotaviruses, amino acid identity and similarity were highest with human rotaviruses. Additionally, the study found that the present virus had some variations in its amino acid sequences and findings from amino acid sequence identity and similarity are almost compatible with nucleotide sequences. Our results suggested that the present bovine RVA field isolate might be genetic ancestor of canine, equine and feline rotaviruses and support the findings of previous studies about interspecies transmission.

There has been an important change in the clinical characteristics and immune profile of Coronavirus disease 2019 (COVID-19) patients during the pandemic thanks to the extensive vaccination programs. Here, we highlight recent studies on COVID-19, from the clinical and immunological characteristics to the protective and risk factors for severity and mortality of COVID-19. The efficacy of the COVID-19 vaccines and potential allergic reactions after administration are also discussed. The occurrence of new variants of concerns such as Omicron BA.2, BA.4, and BA.5 and the global administration of COVID-19 vaccines have changed the clinical scenario of COVID-19. Multisystem inflammatory syndrome in children (MIS-C) may cause severe and heterogeneous disease but with a lower mortality rate. Perturbations in immunity of T cells, B cells, and mast cells, as well as autoantibodies and metabolic reprogramming may contribute to the long-term symptoms of COVID-19. There is conflicting evidence about whether atopic diseases, such as allergic asthma and rhinitis, are associated with a lower susceptibility and better outcomes of COVID-19. At the beginning of pandemic, the European Academy of Allergy and Clinical Immunology (EAACI) developed guidelines that provided timely information for the management of allergic diseases and preventive measures to reduce transmission in the allergic clinics. The global distribution of COVID-19 vaccines and emerging severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants with reduced pathogenic potential dramatically decreased the morbidity, severity, and mortality of COVID-19. Nevertheless, breakthrough infection remains a challenge for disease control. Hypersensitivity reactions (HSR) to COVID-19 vaccines are low compared to other vaccines, and these were addressed in EAACI statements that provided indications for the management of allergic reactions, including anaphylaxis to COVID-19 vaccines. We have gained a depth knowledge and experience in the over 2 years since the start of the pandemic, and yet a full eradication of SARS-CoV-2 is not on the horizon. Novel strategies are warranted to prevent severe disease in high-risk groups, the development of MIS-C and long COVID-19.

A potential immunological silver bullet for COVID-19: the trivalent chimpanzee adenoviral serotype-68 vector (Tri:ChAd)

The current monkeypox disease (MPX) outbreak constitutes a new threat and challenge for our society. With more than 55,000 confirmed cases in 103 countries, World Health Organization declared the ongoing MPX outbreak a Public Health Emergency of International Concern (PHEIC) on July 23, 2022. The current MPX outbreak is the largest, most widespread, and most serious since the diagnosis of the first case of MPX in 1970 in the Democratic Republic of the Congo (DRC), a country where MPX is an endemic disease. Throughout history, there have only been sporadic and self-limiting outbreaks of MPX outside Africa, with a total of 58 cases described from 2003 to 2021. This figure contrasts with the current outbreak of 2022, in which more than 55,000 cases have been confirmed in just 4 months. MPX is, in most cases, self-limiting, however, severe clinical manifestations and complications have been reported. Complications are usually related to the extent of virus exposure and patient health status, generally affecting children, pregnant women, and immunocompromised patients. The expansive nature of the current outbreak leaves many questions that the scientific community should investigate and answer in order to understand this phenomenon better and prevent new threats in the future. In this review, 50 questions regarding monkeypox virus (MPXV) and the current MPX outbreak were answered in order to provide the most updated scientific information and to explore the potential causes and consequences of this new health threat.

As a zoonotic disease that has already spread globally to several million human beings and possibly to domestic and wild animals, eradication of coronavirus disease 2019 (COVID‐19) appears practically impossible. There is a pressing need to improve our understanding of the immunology of this disease to contain the pandemic by developing vaccines and medicines for the prevention and treatment of patients. In this review, we aim to improve our understanding on the immune response and immunopathological changes in patients linked to detoriating clinical conditions such as, cytokine storm, acute respiratory distress syndrome, autopsy findings and changes in acute phase reactants and serum biochemistry in COVID‐19. Similar to many other viral infections, asymptomatic disease is present in a significant but currently unknown fraction of the affected individuals.In the majority of the patients, a one‐week, self‐limiting viral respiratory disease typically occurs, which ends with the development of neutralizing anti‐viral T cell and antibody immunity. The IgM, IgA and IgG type virus‐specific antibodies levels are important measurements to predict population immunity against this disease and whether cross‐reactivity with other coronaviruses is taking place.High viral‐load during the first infection and repeated exposure to virus especially in healthcare workers can be an important factor for severity of disease. It should be noted that many aspects of severe patients are unique to COVID‐19 and are rarely observed in other respiratory viral infections, such as severe lymphopenia and eosinopenia, extensive pneumonia and lung tissue damage, a cytokine storm leading to acute respiratory distress syndrome and multiorgan failure. Lymphopenia causes a defect in antiviral and immune regulatory immunity. At the same time, a cytokine storm starts with extensive activation of cytokine‐secreting cells with innate and adaptive immune mechanisms both of with contribute to a poor prognosis. Elevated levels of acute phase reactants and lymphopenia are early predictors of high disease severity. Prevention of development to severe disease, cytokine storm, acute respiratory distress syndrome and novel approachs to prevent their development will be main routes for future research areas. As we learn to live amidst the virus, understanding the immunology of the disease can assist in containing the pandemic and in developing vaccines and medicines to prevent and treat individual patients.

Sığır koronavirüs (BCoV) ve sığır rotavirüs (BRV) enfeksiyonları, sığır yetiştiriciliğindeki önemli sorunlardan biri olan neonatal buzağı ishallerinin en sık rastlanan viral etkenleri arasında yer almaktadır. Bu çalışmada, Kırıkkale ve çevre illerindeki neonatal ishalli 110 buzağıda BCoV ve BRV etkenleri RT-PCR ile araştırıldı ve pozitif BRV örnekleri PCR ile VP4 ve VP7 genleri temelli olarak genotiplendirildi. RT-PCR sonuçlarına göre toplamda 41 örnek BCoV pozitif (%37,27), 41 örnek BRV pozitif (%37,27) ve 20 örnek hem BCoV hem de BRV pozitif (%18,18) olarak belirlendi. Bu sonuçlara göre Kırıkkale, Kırşehir, Çankırı, Çorum, Yozgat ve Ankara illerinde yetiştirilen buzağılarda BCoV ve BRV, neonatal buzağı ishallerinin etkenleri olarak tespit edildi. BRV genotiplendirme sonuçlarına göre ise bu illerde ülkemizde yaygın olarak görülen genotipler olan G6P[5], G10P[5], G10P[11] ve G6P[11] genotiplerinin sirküle olduğu belirlendi. Bu genotiplerin buzağılarda saptanması, neonatal ishallere karşı aşılamanın ve aşılarda kullanılacak olan suş seçiminin önemini göstermektedir., Bovine coronavirus (BCoV) and bovine rotavirus (BRV) infections are very common in neonatal diarrhoea of calves which are of the important problems in cattle breeding. In the present study 110 calves with neonatal diarrhoea from Kirikkale and surrounding provinces were investigated for BCoV and BRV presence by RT-PCR in stool samples and positive BRV samples were genotyped by PCR based on VP4 and VP7 genes. In total, 41 samples were BCoV positive (37.27%) and 41 samples were BRV positive (37.27%), whereas 20 samples were positive for both BCoV and BRV (18.18%). According to the results of the study, BCoV and BRV are the neonatal calf diarrhoea agents in calves reared in Kirikkale, Kirsehir, Ankara, Cankiri, Corum, and Yozgat provinces. Genotyping results of positive BRV samples indicated that G6P[5], G10P[5], G10P[11], and G6P[11] genotypes, commonly seen in Turkey, are circulating among these provinces. Detection of these genotypes indicated importance of vaccination against neonatal diarrhoea and selection of vaccine strain.

Background Since the first reports of coronavirus disease 2019 (COVID‐19) in Wuhan, China, in December 2019, there have been 198 million confirmed cases worldwide as of August 2021. The scientific community has joined efforts to gain knowledge of the newly emerged virus named severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2), the immunopathological mechanisms leading to COVID‐19, and its significance for patients with allergies and asthma. Methods Based on the current literature, recent advances and developments in COVID‐19 in the context of allergic diseases were reviewed. Results and Conclusions In this review, we discuss the prevalence of COVID‐19 in subjects with asthma, attacks of hereditary angioedema, and other allergic diseases during COVID‐19. Underlying mechanisms suggest a protective role of allergy in COVID‐19, involving eosinophilia, SARS‐CoV‐2 receptors expression, interferon responses, and other immunological events, but further studies are needed to fully understand those associations. There has been significant progress in disease evaluation and management of COVID‐19, and allergy care should continue during the COVID‐19 pandemic. The European Academy of Allergy & Clinical Immunology (EAACI) launched a series of statements and position papers providing recommendations on the organization of the allergy clinic, handling of allergen immunotherapy, asthma, drug hypersensitivity, allergic rhinitis, and other allergic diseases. Treatment of allergies using biologics during the COVID‐19 pandemic has also been discussed. Allergic reactions to the COVID‐19 vaccines, including severe anaphylaxis, have been reported. Vaccination is a prophylactic strategy that can lead to a significant reduction in the mortality and morbidity associated with SARS‐CoV‐2 infection, and in this review, we discuss the proposed culprit components causing rare adverse reactions and recommendations to mitigate the risk of anaphylactic events during the administration of the vaccines.

The coronavirus disease 2019 pandemic (COVID-19), caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has caused an unprecedented global social and economic impact, and numerous deaths. Many risk factors have been identified in the progression of COVID-19 into a severe and critical stage, including old age, male gender, underlying comorbidities such as hypertension, diabetes, obesity, chronic lung disease, heart, liver and kidney diseases, tumors, clinically apparent immunodeficiencies, local immunodeficiencies, such as early type-I interferon secretion capacity, and pregnancy. Possible complications include acute respiratory distress syndrome, shock, disseminated coagulopathy, acute kidney injury, pulmonary embolism, and secondary bacterial pneumonia. The development of lymphopenia and eosinopenia are laboratory indicators of COVID-19. Laboratory parameters to monitor disease progression include lactate dehydrogenase, procalcitonin, high-sensitivity C-reactive protein, proinflammatory cytokines such as interleukin (IL)-6, IL-1, Krebs von den Lungen-6 (KL-6) and ferritin. The development of a cytokine storm and extensive chest computed tomography imaging patterns are indicators of a severe disease. In addition, socioeconomic status, diet, lifestyle, geographical differences, ethnicity, exposed viral load, day of initiation of treatment, and quality of health care have been reported to influence individual outcomes. In this review, we highlight the scientific evidence on the risk factors of COVID-19.

In December 2019, China reported the first cases of the coronavirus disease 2019 (COVID-19). This disease, caused by the severe acute respiratory syndrome-related coronavirus 2 (SARS-CoV-2), has developed into a pandemic. To date it has resulted in ~5.6 million confirmed cases and caused 353,334 related deaths worldwide. Unequivocally, the COVID-19 pandemic is the gravest health and socio-economic crisis of our time. In this context, numerous questions have emerged in demand of basic scientific information and evidence-based medical advice on SARS-CoV-2 and COVID-19. Although the majority of the patients show a very mild, self-limiting viral respiratory disease, many clinical manifestations in severe patients are unique to COVID-19, such as severe lymphopenia and eosinopenia, extensive pneumonia, a “cytokine storm” leading to acute respiratory distress syndrome, endothelitis, thrombo-embolic complications and multiorgan failure. The epidemiologic features of COVID-19 are distinctive and have changed throughout the pandemic. Vaccine and drug development studies and clinical trials are rapidly growing at an unprecedented speed. However, basic and clinical research on COVID-19-related topics should be based on more coordinated high-quality studies. This paper answers pressing questions, formulated by young clinicians and scientists, on SARS-CoV-2, COVID-19 and allergy, focusing on the following topics: virology, immunology, diagnosis, management of patients with allergic disease and asthma, treatment, clinical trials, drug discovery, vaccine development and epidemiology. Over 140 questions were answered by experts in the field providing a comprehensive and practical overview of COVID-19 and allergic disease.

Bu calismanin amaci Schmallenberg virus (SBV) seropozitif belirlenen klinik olarak saglikli Akkaraman koyunlarda bazi oksidatif stres parametrelerinin (nitrik oksit (NO), malondialdehid (MDA), total antioksidan kapasite (TAK), total oksidan kapasite (TOK), katalaz aktivitesi) degerlendirilmesiyle oksidan-antioksidan durumun belirlenmesidir. Calismanin materyalini, ELISA yontemiyle SBV ozgul antikorlari yonunden 4 pozitif, 6 supheli ve 7 negatif-kontrol kan serumu olusturdu. Gruplar arasinda istatistiksel bir fark olmamasina ragmen (P>0.05) en yuksek NO seviyesi SBV seropozitif serumlarindayken, en dusuk TOK seviyesi SBV negatif serumlarda bulundu. Gruplardaki TAS degerleri oldukca benzerdi (P=0.981). SBV seropozitif ve supheli serum MDA seviyeleri benzerken SBV seropozitif ve negatif serumlarin MDA seviyeleri istatistiki olarak farkli bulundu (P=0.021). SBV seropozitif olan serumlarin katalaz aktiviteleri, SBV supheli serumlarin katalaz aktivitelerinden daha dusuk (P=0.030) iken pozitif ve negatif gruplarin katalaz aktiviteleri ile supheli ve negatif gruplarin katalaz aktiviteleri benzer bulundu. Sonuc olarak; hastaliga ait klinik semptom bulunmayan SBV seropozitif Akkaraman irki hayvanlarda MDA seviyesinin etkilendigi belirlendi.

In December 2019, China reported the first cases of the coronavirus disease 2019 (COVID‐19). This disease, caused by the severe acute respiratory syndrome–related coronavirus 2 (SARS‐CoV‐2), has developed into a pandemic. To date, it has resulted in ~9 million confirmed cases and caused almost 500 000 related deaths worldwide. Unequivocally, the COVID‐19 pandemic is the gravest health and socioeconomic crisis of our time. In this context, numerous questions have emerged in demand of basic scientific information and evidence‐based medical advice on SARS‐CoV‐2 and COVID‐19. Although the majority of the patients show a very mild, self‐limiting viral respiratory disease, many clinical manifestations in severe patients are unique to COVID‐19, such as severe lymphopenia and eosinopenia, extensive pneumonia, a “cytokine storm” leading to acute respiratory distress syndrome, endothelitis, thromboembolic complications, and multiorgan failure. The epidemiologic features of COVID‐19 are distinctive and have changed throughout the pandemic. Vaccine and drug development studies and clinical trials are rapidly growing at an unprecedented speed. However, basic and clinical research on COVID‐19–related topics should be based on more coordinated high‐quality studies. This paper answers pressing questions, formulated by young clinicians and scientists, on SARS‐CoV‐2, COVID‐19, and allergy, focusing on the following topics: virology, immunology, diagnosis, management of patients with allergic disease and asthma, treatment, clinical trials, drug discovery, vaccine development, and epidemiology. A total of 150 questions were answered by experts in the field providing a comprehensive and practical overview of COVID‐19 and allergic disease., NHMRC Senior Research Fellowship, Grant/ Award Number: GNT1117687; Consejo Superior de Investigaciones Científicas, Grant/Award Number: IJCI-2016-27619; Instituto de Salud Carlos III, Grant/Award Number: JR19/0029

WOS:000556848300001 PubMed: 32757829 Schmallenberg virus (SBV), discovered in Germany in 2011, causes congenital malformations in ruminants. Reverse-transcription real-time PCR (RT-rtPCR) assays based on various segments of SBV have been developed for molecular detection. We developed alternative RT-rtPCR assays for SBV detection to avoid earlier reported mutations and hypervariable regions of the S and M segments of the viral genome. For SYBR Green-based detection of the S segment, theR(2)value and efficiency of the developed assay were 0.99 and 99%, respectively. For probe-based S segment detection, 2 assays were developed; the first had anR(2)value of 0.99 and 102% efficiency, and the second had aR(2)value of 0.98 and 86% efficiency. The probe-based M segment assay had anR(2)value of 1.00 and 103% efficiency. Detection limits of the RT-rtPCR assays with new primer sets were 10(2)and 10(1)copies/mu L for the S and M segments, respectively. Field samples from cattle and sheep were also used for primary validation of the developed assays. Our assays should be suitable for SBV detection in ruminants and for in vitro studies of various SBV strains. General Directorate of Agricultural Research and Policies, Ministry of Agriculture and Forestry, Republic of Turkey [TAGEM15/AR-GE48] This study is a part of a project supported by the General Directorate of Agricultural Research and Policies, Ministry of Agriculture and Forestry, Republic of Turkey (grant TAGEM15/AR-GE48).

Sursal, Neslihan/0000-0002-4144-9520; Ozen, Dogukan/0000-0003-1943-2690; YILDIZ, Kader/0000-0001-5802-6156; WOS: 000456711400001 Toxoplasma (T.) gondii is an obligatory intracellular apicomplexan parasite. The donkey is one of the intermediate hosts of T. gondii. There are almost no data about T. gondii infection in donkeys, apart from a few seroprevalence reports. The aim of the study was to detect the formation of extracellular traps by donkey polymorphonuclear neutrophils (PMNs) when exposed to T. gondii tachyzoites in vitro. Tachyzoites were observed to be entrapped within extracellular traps (NET) structures. Myeloperoxidase, neutrophil elastase (NE), and histone (H3) were observed in NET structures. NE and reactive oxygen species activity increased with time and was higher in the PMN-tachyzoite cocultures. Some tachyzoites were seen close localization to the nucleus of neutrophils. Degranulation and extracellular traps were observed simultaneously in some donkey neutrophils after incubation for 1 hour. The tachyzoite invasion rate decreased in PMN-tachyzoite cocultures in comparison to the controls. NETs can negatively affect the viability of entrapped tachyzoites in addition to their action of physical entrapment. NE may play a key role in the viability of T. gondii in donkeys. The amount of extracellular DNA increased with incubation time in the PMN-tachyzoite cocultures (P < .001). This is a first report regarding the formation of donkeys NETs after exposure to T. gondii tachyzoites in vitro. Unlike sheep and cattle PMN, degranulation and extracellular traps developed in the same neutrophil. Further studies focusing on signaling pathways may shed light on what determines the behavior of donkey neutrophils after exposure to T. gondii. (C) 2018 Elsevier Inc. All rights reserved.

AKSOY, Emel/0000-0001-9536-4322 WOS: 000459994300010 PubMed: 30541984 Schmallenberg virus (SBV), discovered in 2011 in Germany, is associated with clinical manifestations of fever, diarrhea, reduced milk yield, abortions and congenital malformations in ruminants. Despite many studies performed for SBV, there is no detailed research on in vitro apoptotic effect of SBV. This study is aimed to determine apoptosis pathways and role of proapoptotic and anti-apoptotic molecules in Vero cells infected with SBV. The study results showed that SBV induced apoptosis via both extrinsic and intrinsic pathways by activating both caspase-8 and caspase-9, respectively. Expression analyses of pro-apoptotic (Bax, Bak and Puma) and anti-apoptotic (Bcl-2 and Bcl-XL) genes revealed that SBV-induced apoptosis causes upregulation of pro-apoptotic genes, dominantly via Puma gene, whereas Bcl-2 and Bcl-XL genes were downregulated. In conclusion, this is the first detailed report about SBV induced apoptosis in the Vero cells via both extrinsic and intrinsic cascades and apoptosis induction is seem to be regulated by Puma. Scientific Research Project Coordination Unit (BAP) of Kirikkale University [KU-BAP 2015/052] This work was supported by the Scientific Research Project Coordination Unit (BAP) of Kirikkale University [grant number KU-BAP 2015/052] and includes part of PhD dissertation of Emel AKSOY. The authors would also like to thank Prof. Dr. Latif OZTURK (Kirikkale University, Faculty of Economics and Administrative Sciences, Department of Econometrics, Kirikkale, Turkey) for his advice and help in statistical analysis.

The aim of the study was to determine the epidemiological data of bovine viral diarrhea virus (BVDV), bovine herpesvirus-1 (BHV-1), bovine herpesvirus-4 (BHV-4), bovine herpesvirus-5 (BHV-5) and Brucella-associated cattle that were previously reported to have abortion and infertility problems in Ankara, Corum, Kirikkale and Yozgat provinces, Turkey. Whole blood and sera samples were obtained from 656 cattle, and antibodies against Brucella spp. were detected in 45 (6.86%) and 41 (6.25%) animals by Rose Bengal plate and serum tube agglutination tests, respectively. The seropositivity rates against BVDV, BHV-1 and BHV-4 were 70.89%, 41.3% and 28.78%, respectively. RT-PCR and PCR were performed to detect RNA and DNA viruses in blood samples, respectively. The BVDV 5'-untranslated region and BHV-1 gB gene detected in this study were phylogenetically analyzed. The BVDV strains analyzed in this study were closely related to those previously reported from Turkey. The nucleotide sequence from the BHV-1 strain detected in this study is the first nucleotide sequence of BHV-1 circulating in this area of Turkey deposited in the GenBank. The presence of Brucella spp. and prevalence of BHV-1, BHV-4 and BVDV in cattle should be further investigated throughout these regions.

abayli, hasan/0000-0003-2116-105X; WOS: 000410909200039 PubMed: 28674863 Relatively high prevalence and mortality rates of bovine ephemeral fever (BEF) have been reported in recent epidemics in some countries, including Turkey, when compared with previous outbreaks. A limited number of complete genome sequences of BEF virus (BEFV) are available in the GenBank Database. In this study, the complete genome of highly pathogenic BEFV isolated during an outbreak in Turkey in 2012 was analyzed for genetic characterization. The complete genome of the Turkish BEFV isolate was amplified by reverse transcription-polymerase chain reaction (RT-PCR) and sequenced. It was found that the complete genome of the Turkish BEFV isolate was 14,901 nt in length. The complete genome sequence obtained from the study showed 91-92% identity at nucleotide level to Australian (BB7721) and Chinese (Bovine/China/Henan1/2012) BEFV isolates. Phylogenetic analysis of the glycoprotein gene of the Turkish BEFV isolate also showed that Turkish isolates were closely related to Israeli isolates. Because of the limited number of complete BEFV genome sequences, the results from this study will be useful for understanding the global molecular epidemiology and geodynamics of BEF. Scientific and Technological Research Council of Turkey (TUBITAK)Turkiye Bilimsel ve Teknolojik Arastirma Kurumu (TUBITAK) [114O668] This research was financially supported by the Scientific and Technological Research Council of Turkey (TUBITAK Project no. 114O668).

YILDIZ, Kader/0000-0001-5802-6156; Sursal, Neslihan/0000-0002-4144-9520; Babur, Cahit/0000-0002-6524-3260 WOS: 000406567800001 PubMed: 28669381 The main aim of this study was to compare extracellular traps (NETs) formation by polymorphonuclear neutrophils (PMNs) of cattle and sheep when exposed to T. gondii tachyzoites in vitro. The effects of parasite concentrations and different incubation periods on NETs development in cattle and sheep PMNs were studied. The effect of NET structures on host cell invasion by tachyzoites was also studied. This is the first report of NETs development by sheep and cattle PMNs against T. gondii in vitro. T. gondii-induced extracellular DNA production from PMNs was dependent on tachyzoite concentrations and incubation time in both sheep and cattle. Many nuclear and cytoplasmic changes were observed in sheep and cattle PMNs after exposure to T. gondii tachyzoites. The typical appearance of NETs, with MPO, NE and histone (H3) attached to extracellular DNA, was observed. Tachyzoites were entrapped within this structure. Myeloperoxidase (MPO) activity was higher in the cattle PMNtachyzoite co-cultures than sheep. NETs structures released from sheep PMNs caused mechanical immobilisation of T. gondii tachyzoites, however, NET structures released from cattle PMNs may be lethal to tachyzoites. Bovine MPO may have a lethal effect on T. gondii tachyzoites in vitro during a 3 h incubation. Besides other mechanisms that effect on host susceptibility to T. gondii in sheep and cattle, extracellular traps formation as a part of immunological reactions may be play a role in host susceptibility to T. gondii. Scientific and Technological Research Council of Turkey (TUBITAK TOVAG)Turkiye Bilimsel ve Teknolojik Arastirma Kurumu (TUBITAK) [214O288] We would like to thank Prof. Dr. Serdar Diker from Department of Microbiology of Ankara University Faculty of Veterinary Medicine for his excellent technical support during fluorometric analysis. We are also deeply grateful to Dr. Dogukan Ozen from Biostatistic Department of Ankara University Faculty of Veterinary Medicine for his excellent support in statistical analysis. This work was financially supported by The Scientific and Technological Research Council of Turkey (TUBITAK TOVAG, project no: 214O288). Portion of data was presented at 5th European Immunology & Innate Immunity, Berlin, Germany, July 21-23, 2016.

Avrupa'da 2011 yılında ruminantları etkileyen yeni bir virüs belirlenmiştir. Schmallenberg olarak isimlendirilen bu virüsün Türkiye'deki yaygınlığı üzerine yapılan çalışmalar oldukça sınırlı sayıdadır. Bu çalışmada; koyun yetiştiriciliğinde büyük ekonomik kayıplara neden olan Schmallenberg virüsün (SBV) seroprevalansı ve Culicoides'lerle nakledilmesinden dolayı bazı coğrafi özellikler (rakım, büyük su kaynaklarına yakın olması) ile seroprevalans arası ilişkinin araştırılması amaçlanmıştır. Bu amaçla, Kırıkkale merkez ve ilçelerinde, 684 ile 1219 m arası rakımda bulunan, 38 koyun sürüsündeki 1038 hayvandan (969 koyun, 69 koç) kan serum örneği alınarak ticari ELISA kiti ile SBV özgül antikorları yönünden değerlendirilmiştir. Çalışmaya alınan hayvanların yaşları 2 - 4 (n517), 4 - 6 (n474), 6 (n47) olarak gruplandırılmıştır. Örneklenen hayvanların %0.38'i (4/1038) pozitif, %0.57'si (6/1038) şüpheli olarak değerlendirilmiştir. SBV antikorları yönünden pozitif olarak belirlenen dört hayvanın birinin koç, üçünün koyun; şüpheli olarak değerlendirilen altı hayvanın ise birinin koç, beşinin koyun olduğu belirlenmiştir. Pozitif ve şüpheli olarak belirlenen hayvanların bulunduğu sürülerin belli bir rakımda yoğunlaşmadığı ve bu sürülerin yarısının büyük su kaynaklarına yakın olmadığı tespit edilmiştir. SBV yönünden pozitif örnek sayısı az olduğu için seroprevalansı ile yaş grupları, cinsiyet ve bazı coğrafi özellikler arasındaki ilişki istatistiksel olarak değerlendirilmemiştir. Kırıkkale'de yetiştirilen koyunlarda, SBV seroprevalansının araştırıldığı bu çalışma ile enfeksiyonun varlığı ilk kez ortaya konulmuş olup, abortus/anomalili yavru doğumları gözlenen sürülerde, söz konusu enfeksiyonun da araştırılmasının uygun olabileceği kanısına varılmıştır. A virus, which effects ruminants, was firstly described in 2011 in Europe. There are limited information about this virus called Schmallenberg in Turkey. The aim of the present study was to determine the seroprevalance of Schmallenberg virus (SBV), which may cause major economic losses to sheep, and the association between seroprevalance and some geographical features (e.g. altitude, near a large water source) because of transmission by Culicoides spp. Blood samples from 1038 animals (969 ewes, 69 rams) in 38 flocks were collected and evaluated with a commercial ELISA kit in terms of specific antibody for SBV in centrum and eight towns of Kirikkale, where the altitudes ranges from 684 to 1219 meters. Age groups of the animals were determined as >2 - 4 - 6 (n 47) years old. The samples of Schmallenberg virus specific antibody detected as positive and suspected were 0.38% (4/1038) and 0.57% (6/1038), respectively. One out of four Schmallenberg virus specific antibody positive animals was ram, the others were ewes. On the other hand, one out of six Schmallenberg virus specific antibody suspected animals was ram, the others were ewes. The flocks of the animals with the virus specific antibody positive and suspected animals were not in places with the certain altitudes and the half of those flocks were not close to large water sources. Due to the limited numbers of animals with the virus specific antibody positive, no statistical analysis were performed to see the association between the seroprevalance of the virus and gender, age of the animals and the geographical features of the places. This study that is first investigation about Schmallenberg virus seroprevalence in sheep in Kirikkale, showed the presence of the virus specific antibody. It was also thought that this virus should be considered as the cause of abortion and malformed births in flocks.

Herpes simplex virus (HSV) infection of the cornea culminates in an immunopathological lesion (stromal keratitis--SK) that impairs vision. This report shows that HSV infection results in IL-23 up-regulation, but if this response fails to occur, as was noted in p19-/- mice, the severity of lesions, their incidence and the level of viral induced angiogenesis were significantly increased compared to wild-type (WT) animals (p

abayli, hasan/0000-0003-2116-105X; WOS: 000369159000008 Schmallenberg virus (SBV) infection emerged in European domestic and wild ruminants in 2011. There is very limited information about the characterization of SBV isolates and the epidemiology of its infections in the rest of world, except for in European countries. We investigated the circulation of SBV in cattle herds in Central Anatolia, Turkey, in 2013. A total of 180 whole-blood samples were analyzed using real-time RT-PCR. The presence of SBV RNA was detected in 6 (3.3%) samples. For phylogenetic analysis and confirmation of real-time RT-PCR results, the S gene segment was amplified, sequenced, and compared to other segments. In addition, SBV-specific antibodies were detected in 87 (24.1%) of 360 sera using a virus neutralization test. In the S gene sequence analysis of four randomly selected samples, 98%-99% nucleotide identity was observed between our strains and SBVs isolated in European countries between 2011 and 2013. The results of this study indicate that SBV was in Turkey in 2013. Furthermore, the sequencing results suggest that it could be the same virus that is in European countries.

sozdutmaz, ibrahim/0000-0002-3712-7991; GAZYAGCI, SERKAL/0000-0002-0043-6942 WOS: 000368558800003 Abortion, neonatal deaths, and infertility cause important production losses in the livestock industry. Many viral infections are responsible for changes in hematological values of animals. This study aimed to find whether there is any difference in blood parameters of cattle that are naturally infected with BVDV, BHV-1, and BHV-4 and Brucella-seronegative. For this purpose, 401 whole-blood samples collected from cattle were analyzed by ELISA and PCR/RT-PCR to detect specific antibodies and antigens for BVDV, BHV-1, and BHV-4. Hematological analysis and results were compared between naturally infected BVDV, BHV-1, and BHV-4 groups and a control group. Results showed a statistically significant increase in hematological values of group 2 with BVDV compared to the corresponding parameters of group 1 (control) for mean corpuscular volume (MCV), hematocrit (Hct%), mean corpuscular hemoglobin concentration (MCHC), and mean platelet volume (MPV). The same was seen for hematological values of BHV-1 for MCV, Hct (%), and MPV and of BHV-4 for MPV (P < 0.005). However, it was determined that statistical differences in hematological values of naturally infected animals cannot be used as markers for clinical diagnosis of viral infections. Thus, we suggest that evaluations of hematological parameters with specific diagnostic tests are necessary to obtain more reliable results for the diagnosis of viral diseases. Scientific and Technological Research Council of TurkeyTurkiye Bilimsel ve Teknolojik Arastirma Kurumu (TUBITAK) [TUBITAK-111O871]; KUBAP [2011/35] This study was summarized from a part of the dissertation of ME Aslan and was supported by the Scientific and Technological Research Council of Turkey (TUBITAK-111O871) and KUBAP (2011/35).

After ocular infection, HSV-specific CD8+ T cells migrate to and are specifically retained in the ophthalmic branch of the trigeminal ganglia (TG) even at the time when replicating virus is no longer evident. Virus-specific CD8+ T cells maintain an activation phenotype and secrete IFN-γ in the latent TG. In this report we demonstrated that activated virus-specific memory CD8+ T cells, although potentially cytolytic, also expressed the CD94-NK cell receptor subfamily G2a inhibitory molecule and were unable to exert cytotoxicity when engaged by Qa-1b expressing targets. Interestingly, many neurons in the latent TG expressed Qa-1b, and blocking of Qa-1b/CD94-NKG2a interaction in an ex vivo TG culture resulted in neuronal cell lysis. The expression of the inhibitory CD94-NKG2a molecule could be induced by TGF-β1, which was shown to present as a bioactive molecule in the latent TG. Additionally, CD4+ forkhead/winged helix transcription factor 3+ T cells were also determined in the latent TG. Our results demonstrate the operation of a regulatory system in vivo that serves to protect irreplaceable neurons from destruction by the immune system.

CD4+CD25+ regulatory T cells (Treg) can inhibit a variety of autoimmune and inflammatory diseases, but their involvement in regulating virus-induced immunopathology is not known. We have evaluated the role of Treg in viral immunopathological lesion stromal keratitis. This frequent cause of human blindness results from a T cell-mediated immunoinflammatory response to HSV in the corneal stroma. The results show that lesions were significantly more severe if mice were depleted of Treg before infection. The Treg was also shown to modulate lesion expression induced by adoptive transfer of pathogenic CD4+ T cells in infected SCID recipients. The mechanism of Treg control of stromal keratitis involved suppressed antiviral immunity and impaired expression of the molecule required for T cell migration to lesion sites. Interestingly, Treg isolated from ocular lesions in nondepleted mice showed in vitro inhibitory effects involving IL-10, but were not very effective in established lesions. Our results decipher the in vivo role of Treg in a virus-induced immunopathology and imply that manipulation of regulatory cell function represents a useful approach to control viral-induced immunoinflammatory disease.

The aim of this study is to investigate necrosis and apoptosis in epididymal bull spermatozoa before freezing and after thawing using the flow cytometric method and to compare this with eosin-nigrosin dyeing, which is the conventional method used in assessing of spermatozoa. The testicles from fourteen bulls at local slaughterhouse were used for this study. The proportions of live spermatozoa, total apoptotic, necrotic and early necrotic spermatozoa levels were observed via flow cytometry. Annexin V/PI fluoruscence dyeing was used to investigate the proprotions of apoptotic, necrotic, early necrotic and live spermatozoa for flow cytometry. The proportion of dead spermatozoa and protoplasmic droplets were determined using the eosin-nigrosin conventional dyeing method in fresh and frozen-thawed spermatozoa. The average dead spermatozoa count with flow cytometry was less than with the eosin-nigrosin method (P

BERBER, ENGIN/0000-0003-0420-7714; WOS: 000332437300016 PubMed: 23800972 Regional cases of bovine ephemeral fever (BEF) were documented previously in Turkey. Previous cases were confirmed in South-East Turkey with low cow mortality. Recent BEF-suspected outbreaks with high mortality were documented in many regions of Turkey in 2012. The aim of study was the epidemiological examination of the outbreak and molecular characterization of the viruses detected from the outbreak. For this reason, blood samples were collected from BEF-suspected outbreak regions. From the results of RT-PCR, high rate of BEF-suspected samples (48/60 or 80%) was found positive for BEF virus (BEFV) RNA. The nucleotide sequences of the G(1) region of G gene of BEFV in the current study during the 2012 outbreak were grouped into cluster II of BEFV. It was suggested that BEFV may be spread out to other neighbor countries in the future years.

Ozan, Emre/0000-0001-9872-8152; Risvanli, Ali/0000-0001-5653-0025; Ozenc, Erhan/0000-0002-9179-8796 WOS: 000327431300026 PubMed: 23645515 This, partly retrospective study, was designed to determine the seroprevalence of Schmallenberg virus (SBV), a new Orthobunyavirus first reported in Germany in late 2011, in domestic ruminants from the Middle Black Sea, West, and Southeast regions of Turkey. An indirect enzyme-linked immunosorbent assay was used to screen serum samples collected from slaughterhouse animals between 2006 and 2013. The overall seroprevalence was 335/1,362 (24.5 %) with 325/816 (39.8 %), 5/307 (1.6 %), 3/109 (2.8 %), and 2/130 (1.5 %) recorded in cattle, sheep, goats, and Anatolian water buffalo, respectively. This is the first study to demonstrate the presence of antibodies to SBV in Turkish ruminants; it indicates that cattle are more susceptible to infection than sheep, goats, or buffalo and that exposure of domestic ruminants to SBV in Turkey may have occurred up to 5 years prior to the first recorded outbreak of the disease in 2011.

GAZYAGCI, SERKAL/0000-0002-0043-6942 WOS: 000287097100004 PubMed: 21104124 In this study, we investigated the changes occurring in the activities of determining the biochemical and hematological parameters in persistently infected sheep with border disease virus (BDV) and control sheep. While cholesterol, aspartate aminotransferase, lactate dehydrogenase, high-density lipoprotein, and glucose parameters were found to be statistically different between control and BDV positive groups (p < 0.01), total protein, alkaline phosphotase, creatine kinase, amylase, glucose, and high-density lipoprotein were found to be statistically different between control and persistently infected group (p < 0.01). Interestingly, all groups were shown only mean corpuscular volume parameter was different (p < 0.01). It was found that cholesterol, aspartate aminotransferase, amylase, high-density lipoprotein, and low-density lipoprotein parameters were different between PI and infected sheep (p < 0.01). It was speculated that BDV might effect also the expression of low-density lipoprotein receptor and determination of the changes in BD and its clinical importance might contribute to the veterinarians and scientists studying in this area.

GAZYAGCI, SERKAL/0000-0002-0043-6942 WOS: 000297671800017 The maedi-visna virus (MVV) classified as a lentivirus of the retroviridae family, causes a very common economically important disease in sheep, in many parts of the worlds. Presences of the infection in Turkey have been shown by researches in previous studies. In this study all blood samples were examined by ELISA and PCR to detect MVV antibody and antigen responses, respectively. Hematological findings were monitored and comparing antibody and antigen positive naturally infected (n=5), antibody positive and antigen negative (n=20), antibody and antigen negative (n=20) in sheep was done. When infected sheep were compared with control sheep, Hgb and MPV parameters were shown statistically different (P

Palomares, Oscar/0000-0003-4516-0369 WOS: 000278343500001 PubMed: 20148422 Allergy is a Th2-mediated disease that involves the formation of specific IgE antibodies against innocuous environmental substances. The prevalence of allergic diseases has dramatically increased over the past decades, affecting up to 30% of the population in industrialized countries. The understanding of mechanisms underlying allergic diseases as well as those operating in non-allergic healthy responses and allergen-specific immunotherapy has experienced exciting advances over the past 15 years. Studies in healthy non-atopic individuals and several clinical trials of allergen-specific immunotherapy have demonstrated that the induction of a tolerant state in peripheral T cells represent a key step in healthy immune responses to allergens. Both naturally occurring thymus-derived CD4(+)CD25(+)FOXP3(+) Treg and inducible type 1 Treg inhibit the development of allergy via several mechanisms, including suppression of other effector Th1, Th2, Th17 cells; suppression of eosinophils, mast cells and basophils; Ab isotype change from IgE to IgG4; suppression of inflammatory DC; and suppression of inflammatory cell migration to tissues. The identification of the molecules involved in these processes will contribute to the development of more efficient and safer treatment modalities. Swiss National FoundationSwiss National Science Foundation (SNSF) [32-125249, 32-118226]; Global Allergy and Asthma European Network; Christine Kuhne-Center for Allergy Research and Education (CK-CARE); Ministerio de Educacion y CienciaSpanish Government; FECYT (Spain) The M. Akdis and C.A. Akdis laboratories' are supported by the Swiss National Foundation grants 32-125249 and 32-118226, Global Allergy and Asthma European Network (GA2LEN), and Christine Kuhne-Center for Allergy Research and Education (CK-CARE). We thank Ministerio de Educacion y Ciencia and FECYT (Spain) for a postdoctoral fellowship to O. Palomares.

Viral infections cause important problems and significant economic loses in sheep and goats that can be protected by an investigation of infections. This study aimed to determine the sheep and goat viral infections including bluetongue virus (BTV), maedivisna virus (MVV) and caprine arthritis-encephalitis virus (CAEV) in Kirikkale city located in Central Anatolia region of Turkey. Blood samples collected from 279 sheep and 146 goats were tested by ELISA, RT-nested PCR and nested PCR. It was detected that antibody response to BTV (49.8%), CAEV (7.5%) and MVV (19.4%) were seropositivity values in all serum samples. BTV (2.87%) and MVV (9.25%) antigens were detected in seropositive whole blood samples via RT-nested PCR and nested PCR but not CAEV. It was found that double positive animals (14.6%) for MVV and BTV in sheep but not goat. As a conclusion sheep and goat were infected with naturally BTV, CAEV and MVV in Kirikkale region. This study showed that this the first report for BTV, MVV and CAEV infections from Kirikkale region. Therefore infections are needed to be further investigations to determine detailed survey studies.

Crimean-Congo hemorrhagic fever virus (CCHFV) is a tick-borne virus in the family Bunyaviridae , genus Nairovirus . The virus is transmitted to humans through infected tick bites or from direct contact with viremic animals or humans. In the present study, a total of 1,015 adult ticks were collected from cattle (603 specimens), sheep (17 specimens), and goats (395 specimens) in the Kelkit Valley in Turkey. Four tick species were recognized on the animals in the surveyed region. The most abundant species were Rhipicephalus bursa and Hyalomma marginatum marginatum , at 47.68% (484/1,015) and 46.40% (471/1,015), respectively. Reverse transcriptase PCR was used to recover partial sequences of the CCHFV small (S) genome segment. The presence of CCHFV was determined in 3 of 33 (9.09%) R. bursa pools and in 1 of 31 (3.22%) H. m. marginatum pools. Virus sequences from R. bursa were extremely different from those of the Greek CCHFV strain (U04958) isolated from an R. bursa tick. Phylogenetic analysis indicated that the CCHFV isolates obtained in this study clustered in group 5, whose range encompasses southwestern Russian and Kosovo. This is the first evidence of CCHFV in ticks from Turkey. Even though Hyalomma is the main vector for CCHFV, R. bursa may play a role in CCHFV transmission.

In this study, we assessed the efficacy of the specific knockdown of matrix metalloprotein-9 (MMP-9) in vitro and in vivo using short hairpin RNA (shRNA) against MMP-9. Two plasmids were generated encoding shRNA (pshRNA) targeted against two distinct MMP-9 gene sequences. Transfection of these pshMMP-9s could be shown to specifically inhibit MMP-9 expression both in vivo and in vitro. The effect occurred in vitro both in cells that endogenously produce MMP-9 and in cells exogenously transfected with an MMP-9-encoding plasmid. Using an in vivo transfection approach, the pshMMP- 9 was also effective at inhibiting MMP-9 protein expression in the mouse cornea. pshMMP-9s were also tested against herpes simplex virus (HSV) in the cornea. Delivery of the pshMMP-9 stopped angiogenesis and decreased the severity of herpetic keratitis. However, interferon-alpha (IFN-alpha) and IFN- beta induced by pshRNA might also contribute to inhibition of herpetic simplex keratitis (HSK) in the cornea.

This report seeks a means of maximizing memory CD8 T cell responses to peptide immunization. Delivery of the CD8 peptide epitope by stress protein, heat shock protein (hsp)70, results in excellent immunogenicity at the acute phase but memory responses were poor both in terms of the number of responding cells as well as their functional avidity. We demonstrate for the first time that hsp70 can also be used as a vehicle to achieve CD4 T cell responses to loaded peptide epitopes and that coimmunization with hsp70 loaded with both CD8 and CD4 peptide epitopes may increase memory up to 3-fold. Furthermore, CD8+ T cell memory responses were of higher avidity measured both by in vitro cytotoxicity assays and a new methodology that measures the avidity of CTL activity in vivo in mice. Our results emphasize that peptide immunization remains a viable approach to induce long-term CD8+ T cell function, providing steps are taken to assure appropriate stimulation of Th cell responses.