Diagnosis and phylogenetic analysis of bovine papillomaviruses in cattle papillomatosis cases by different methods.

Papillomaviruses, known as epitheliotropic, cause proliferation in the skin, mucosa, and different visceral organs. In this study, it was aimed to diagnose bovine papillomavirus (BPV) by using different methods in the lesion taken from twenty cattle with papillomas in different areas of the body and to reveal its molecular characterization. In our study, molecular, immunohistochemistry, and transmission electron microscopy (TEM) methods were used for virus identification. Additionally, sequencing analysis was used to ascertain the phylogenetic relationship between the obtained field strains and other isolates submitted to GenBank. Histopathological analyses of the collected samples were done in addition to diagnostic procedures. Intranuclear virus particles were detected when the papillomas were investigated with TEM. In PCR analyses using degenerate and type-specific primer sets, the presence of BPV nucleic acid was determined in 70% (14/20) and 90% (18/20) of the samples, respectively. No virus could be detected in PCR applications using MY 09/11 degenerate primer sets. Twenty animals of different ages, races, and genders included in the study by random sampling method from different herds were divided into 4 groups according to the body regions where the lesions were located. Sequence analysis was performed on a sample from each group that showed strong positivity in the PCR technique using FAP 59/64 degenerate primer set and type-specific primer set. Sequence analyses were performed using FAP 59/64 degenerate primers of amplicons for phylogenetic research. In these analyses, three of the isolated strains were identified as BPV-1, which is in the Deltapapillomavirus 4 genus, and one as BPV-2. As a result of the study, it was concluded that molecular and phylogenetic studies using type-specific primers are more beneficial in order to fully reveal the etiology of papillomatosis in cattle and it would be correct to determine BPV types before prophylactic (vaccine, etc.) applications.

Evaluating the Efficiency of Various Treatment Methods in Cattle Cutaneous Papillomatosis.

In this study, we compared the effectiveness of various methods used in the treatment of cattle with cutaneous papillomatosis. Ivermectin, Tarantula cubensis extract, levamisole, autovaccine, and a combination of T. cubensis extract + levamisole were administered to the animals. The animals were divided into six equal groups. Animals in the control group (n = 10) did not receive any treatment. The animals in the experimental group were administered Ivermectin [three times a week, n = 10, subcutaneous, (SC)], Tarantula cubensis extract (twice a week, n = 10, SC), autologous vaccine (three times at 10-day intervals, n = 10, SC), levamisole [twice at one-week intervals, n = 10, intramuscular (IM)], and levamisole + Tarantula cubensis extract (concurrently). All animals used in the study were monitored for three months at an interval of 15 days. No regression was detected in the papillomas of the control group animals, but recovery was recorded in animals treated with ivermectin at a rate of 70% (7/10), while it was 60% (6/10) in those treated with T. cubensis extract, 100% (10/10) in those treated with autovaccine, 50% (5/10) in those treated with levamisole, and 90% (9/10) in those treated with the combination of T. cubensis extract + levamisole. Significant differences were found between the control group and all treatment groups. Recovery mostly occurred within 45-60 days (P < 0.05). The five treatment modalities applied for the treatment of bovine cutaneous papillomatosis were statistically evaluated and all methods of treatment were effective at different rates. The most precise and effective treatment method was the autovaccine one.

Phylogenetic analysis and searching bovine papillomaviruses in teat papillomatosis cases in cattle by performing histopathology, immunohistochemistry, and transmission electron microscopy.

Papillomaviruses are epitheliotropic in nature and cause proliferation in the skin, mucosa, and various internal organs of various animal species. The lesions they cause, specifically in cattle teats, lead to significant economic losses in the milk industry. In this study, we identified the bovine papillomaviruses (BPVs) responsible for teat papillomas in cattle. The tissue damage caused by the virus was examined histopathologically using immunohistochemical, transmission electron microscopy (TEM), and molecular methods. Additionally, sequence analyses were performed on the isolated field strains to better understand their genetic and phylogenetic relationships with previously reported isolates. Teat papillomatosis was confirmed in the collected samples by histopathological and immunohistochemical methods, which were followed by other diagnostic methods. Intranuclear virus particles were found in the epithelial cells during a TEM examination of teat lesions. BPV was detected in seven samples by performing PCR using degenerate primers and specific primers. The positive samples were used for typing through sequence analysis/PCR with type-specific primers. Three isolates from teat tissues with BPV infection were identified as BPV-6, two as BPV-10, one as BPV-2, and one as BPV-8. The five isolates identified through sequence analysis of positive samples belonged to the Xipapillomavirus 1 genus (one), the Epsilonpapillomavirus 1 genus (one), and the Deltapapillomavirus genus (one) (three). Furthermore, type-specific primers were found to be useful for molecular diagnosis of BPV, which occurs in the etiology of teat papillomas, followed by genotyping and primer generation during characterization. The detection of BPV types and their prevalence, biosafety measures in animal breeding, and the importance of vaccine research are all important.

First molecular evidence of border disease virus in wild boars in Turkey.

Molecular studies on viral diseases in wildlife are limited in Turkey. Pestiviruses infect domestic animals such as pig, cattle, sheep, goats and many other wild ungulates. Cross-species transmission of pestiviruses between wildlife and domestic livestock is a subject of recent concern where wild ungulates are in close contact with domestic ruminants. The International Committee on Virus Taxonomy (ICTV) has named the genus Pestivirus, which belongs to the Flaviviridae family, using the format Pestivirus A, Pestivirus B, Pestivirus C, and so on. Pestivirus A-D replaces Bovine viral diarrhea virus-1 (BVDV-1), Bovine viral diarrhea virus-2 (BVDV-2), Classical swine fever virus (CSFV) and Border disease virus (BDV) respectively. During the 2013-2014 hunting season, a total of 40 samples were collected from wild boars (Sus scrofa ferus) in the area of Western Mediterranean Turkey. In the samples, nucleic acids were investigated for pestivirus, Aujeszky's disease virus, Borna disease virus, coronavirus, mastadenovirus and rotavirus. RT-PCR was performed using primary sets to detect specific partial gene region specific to each virus. Sequence analysis was performed on a positive sample. Phylogenetic analysis revealed that the positive sample, TR/Burdur/13/Boar3, belonged to BDV genotype 1 (Pestivirus D). The first molecular findings of BDV in wild boars in Turkey are reported in this study. This study highlights the importance of further research into diseases that might be transmitted from wild boars to ruminants in Turkey.

Investigation of Norovirus genogroups (GI, GII and GIV) in stool of pet dogs with diarrhea / Investigação de genogrupos de norovírus (GI, GII e GIV) nas fezes de cães de estimação com diarréia

In this study, we searched the existence of human norovirus (NoV) GI, GII and GIV in the stool of 128 pet dogs with diarrhea, of different sex, age and breed, in Burdur, Turkey, using Real-Time PCR method. Human NoV GII was found in only 5 of the 128 dog stool samples (3.91%). It was discovered that human NoV existed most in crossbreed, female and aged 24 months or over dogs. These dogs found with human NoV GII were either bought from pet shops, stray dogs or taken as puppy of another pet dog. The sheltering conditions of these dogs were moderate and they were fed with home food residue and dry food. It was also found that most of them were vaccinated and had certain walking sites. The owners of the animals detected with infection generally did not have the habit of washing their hands or changing their clothes before or after caring their pets. We strongly advice that dog owners' personal hygiene, the necessity of changing their clothes during their contact with animals, the environment provided for the dog, the sensitivity in caring, use of strong and effective disinfectant, keeping the dogs away from toilets and sewerage systems, as well as not feeding them with food residues are crucial issues in dogs' care. Owners of the dogs with NoV GII were middle aged or elderly people, male, and there were no children in their houses. As these dogs are treated like the owner's child, it is assumed that they could be transmitted with NoV GII as a result of close interaction with their owner.(AU)

Neste estudo pesquisamos a existência de norovírus humano (NoV) GI, GII e GIV nas fezes de 128 cães com diarréia, de diferentes sexos, idades e raças, em Burdur, Turquia, utilizando o método de PCR em tempo real. NoV GII humano foi encontrado em apenas 5 das 128 amostras de fezes de cães (3,91%). Foi descoberta NoV humana, principalmente em cruzamentos, fêmeas e cães com idade igual ou superior a 24 meses. Os cães encontrados com NoV GII humano foram comprados de lojas de animais, eram vira-latas ou foram tomados como filhotes de outro cão de estimação. As condições de abrigo desses cães eram moderadas. Os cães foram alimentados com restos de comida caseira e comida seca. Verificou-se também que a maioria dos animais foi vacinada e tinham locais adequados para caminhada. Os donos dos animais detectados com infecção geralmente não tinham o hábito de lavar as mãos ou trocar de roupa antes ou depois de cuidar de seus animais de estimação. Aconselhamos que a higiene pessoal dos donos, a necessidade de trocar de roupa durante o contato com animais, o ambiente fornecido para o cão, a sensibilidade no cuidado, o uso de desinfetantes eficazes, manter os cães longe de banheiros e esgotos, assim como evitar alimentá-los com resíduos alimentares, são questões cruciais no cuidado dos cães. Os proprietários dos cães com NoV GII são de meia-idade ou idosos, a maioria do sexo masculino, e não havia crianças em suas casas. Como esses cães são tratados como um filho, presume-se que eles foram infectados com o NoV GII como resultado de uma interação próxima com o proprietário.(AU)

The role of goats as reservoir hosts for bovine herpes virus 1 under field conditions.

Bovine herpesvirus 1 (BoHV1) is the cause of economically significant viral infections in cattle. Respiratory symptoms associated with the infection are known as Infectious Bovine Rhinotracheitis (IBR). Sheep and goats are less sensitive to the infection although their role in inter-species viral transmission under field conditions is subject to controversy. The objective of this study was to investigate seroprevalence of BoHV1 infections in cattle, sheep, and goats raised together for at least a year. Blood serum samples were taken from 226 cattle, 1.053 sheep, and 277 goats from 17 small- to medium-scale farms. BoHV1-specific antibody presence and titers were determined using virus neutralization test. In total, 73 of the 226 cattle (32.3%) were seropositive. The infection was detected in 13 of the 17 farms. Infection rates ranged from 5.8 to 88.8%. Only one of the 1053 sheep (0.09%) was seropositive. However, 58 of the 277 (20.9%) goats were seropositive. Goat samples taken from 8 of the 17 farms were seropositive with infection rates ranging from 17 to 38.9%. Statistical analysis showed a significant correlation in infection rates between cattle and goats but not sheep. These results suggest that goats may be more sensitive to the BHV1 infection than sheep and the role of goats as possible reservoirs for BoHV1 in the control and eradication of BHV1 in cattle should be considered in future studies.

The first serological evidence for Rift Valley fever infection in the camel, goitered gazelle and Anatolian water buffaloes in Turkey.

Rift valley fever (RVF), a vector-borne zoonotic disease, is caused by a phlebovirus (family Bunyaviridae). The virus was initially characterized approximately 80 years ago in Kenya and disseminated to many countries in the continental Africa, Saudi Arabia, and Yemen. The infection has not been reported in Turkey. In this study, blood serum samples collected from camel (Camelus dromedairus), goitered gazelle (Gazella subgutturosa subgutturosa), and buffaloes (Bubalus bubalis linneaus) from 2000 to 2006 were investigated for RVF using C-ELISA. Camel samples (n = 72) were obtained from private small enterprises in Aydin province in theAegean region. Gazella samples (82) were taken from the biggest captive gazelle herd in Sanliurfa province in the southeast Anatolia. Buffalo samples were collected mostly from small private family type farms in Afyon (168), Amasya (80), Samsun (69), Ankara (35), Sivas (21), Tokat (19), Konya (10), and Elazig (8) provinces in the central, north, west, and east Anatolia. All of the gazella samples were negative; whereas, one of the 71 camel samples (1.3%) was positive for RVF-specific antibodies. Buffalos from Sivas, Tokat, Konya, and Elazig provinces were negative. However, 35 of the 410 samples (8.5%) from rural areas in the following four provinces were positive: Amasya (12/80, 15%), Ankara (5/35, 14.2%), Samsun (8/69, 11.5%), and Afyon (10/168, 5.9%). To our knowledge, this is the first report of presence of RVF infection in Turkey.

The serological and virological investigation of canine adenovirus infection on the dogs.

Two types of Canine Adenovirus (CAVs), Canine Adenovirus type 1 (CAV-1), the virus which causes infectious canine hepatitis, and Canine Adenovirus type 2 (CAV-2), which causes canine infectious laryngotracheitis, have been found in dogs. In this study, blood samples taken from 111 dogs, which were admitted to the Internal Medicine Clinic of Selcuk University, Faculty of Veterinary Medicine, with clinical symptoms. Seventy-seven dogs were sampled from Isparta and Burdur dog shelters by random sampling, regardless of the clinical findings. Dogs showed a systemic disease, characterized by fever, diarrhea, vomiting, oculonasal discharge, conjunctivitis, severe moist cough, signs of pulmonary disease and dehydration. Two dogs had corneal opacity and photophobia. In serological studies, 188 serum samples were investigated on the presence of CAV antibodies by ELISA. Total 103 (103/188-54.7%) blood samples were detected to be positive for CAV antibodies by ELISA. However, 85 (85/188-45.2%) blood samples were negative. Blood leukocyte samples from dogs were processed and inoculated onto confluent monolayers of MDCK cells using standard virological techniques. After third passage, cells were examined by direct immunoflourescence test for virus isolation. But positive result was not detected. In conclusion, this study clearly demonstrates the high prevalence of CAV infection in dogs.

Effects of bovine viral diarrhoea virus on the fertility of cows.

The aim of the present study was to determine the possible relationship between bovine viral diarrhoea (BVD) virus infection and the appearance of cervical mucous discharge (CMD) and the reproductive performance of cows in oestrus. For this purpose, CMD from 97 Holstein cows in oestrus was evaluated visually before artificial insemination (AI). Cows in oestrus were inseminated with frozen semen free from BVD virus (BVDV). Blood samples were tested by enzyme-linked immunoassay (ELISA) for antigen (Ag) and antibodies (Ab) of BVDV. The presence of the BVDV genome in cervical mucus samples was tested by reverse transcriptase-polymerase chain reaction (RT-PCR). The presence of BVDV Ab, Ag or genome was not associated with abnormal cervical mucous discharge (A-CMD). However, the presence of BVDV Ag (but not of the BVDV Ab) in blood samples was associated with a lower first service conception rate (FSCR; 27.8 vs. 70.9%; P < 0.01), indicating that BVDV viraemia at the time of AI has a negative effect on the fertility of cows.

Assessment of feasibility to use computer aided texture analysis based tool for parametric images of suspicious lesions in DCE-MR mammography.

Our aim was to analyze the feasibility of computer aided malignant tumor detection using the traditional texture analysis applied on two-compartment-based parameter pseudoimages of dynamic contrast-enhanced magnetic resonance (DCE-MR) breast image data. A major contribution of this research will be the through-plane assessment capability. Texture analysis was performed on two-compartment-based pseudo images of DCE-MRI datasets of breast data of eight subjects. The resulting texture parameter pseudo images were inputted to a feedforward neural network classification system which uses the manual segmentations of a primary radiologist as a gold standard, and each voxel was assigned as malignant or nonmalignant. The classification results were compared with the lesions manually segmented by a second radiologist. Results show that the mean true positive fraction (TPF) and false positive fraction (FPF) performance of the classifier vs. primary radiologist is statistically as good as the mean TPF and FPF performance of the second radiologist vs. primary radiologist with a confidence interval of 95% using a one-sample t-test with α = 0.05. In the experiment implemented on all of the eight subjects, all malignant tumors marked by the primary radiologist were classified to be malignant by the computer classifier. Our results have shown that neural network classification using the textural parameters for automated screening of two-compartment-based parameter pseudo images of DCE-MRI as input data can be a supportive tool for the radiologists in the preassessment stage to show the possible cancerous regions and in the postassessment stage to review the segmentations especially in analyzing complex DCE-MRI cases.

Detection of central nervous system tissue as bovine spongiform encephalopathy specified risk material in traditional Turkish meat products.

BACKGROUND: This study used enzyme-linked immunosorbent assay kits to investigate the presence of central nervous system (CNS) tissue in commercial raw and processed traditional Turkish meat products offered for consumption in various markets. RESULTS: Ninety-six raw traditional Turkish meat products (32 fresh raw beef patties, 32 cig kofta, 32 pastirma) and 64 processed traditional Turkish meat products (32 doner kebabs and 32 fresh processed beef patties) were analysed. CNS tissue was not found in pastirma, doner kebab, or fresh processed beef patty samples. The levels of CNS contamination in fresh raw beef patties were low (0.1% absorbance standard; 3.1%) and moderate (0.2% absorbance standard; 6.2%). The level of contamination in the cig kofta was low (0.1% absorbance standard; 18.8%). CONCLUSION: CNS tissue was present in all raw traditional Turkish meat products except for pastirma.

Bovine viral diarrhea virus (BVDV) infection in relation to fertility in heifers.

In this study, blood serum and leukocyte samples were collected from 400 Holstein heifers, all of which appeared to be healthy. Antibodies (Ab) against bovine viral diarrhea virus (BVDV) were detected in 57 serum samples, and BVDV antigen (Ag) was detected in 38 leukocyte samples. There were statistically important differences between the average first insemination ages (FIT) of the BVDV (Ag-/Ab+) heifers (p<0.0001) (pregnant p<0.05, nonpregnant p<0.0001) and BVDV (Ag-/Ab-) heifers. The average conception rates (CR) of BVDV (Ag-/Ab+) heifers and BVDV (Ag-/Ab-) heifers were not significant statistically. There were statistically important differences in average FIT between persistent infected (PI) BVDV (Ag+/Ab-) heifers (p<0.0001; PI pregnant p<0.05, PI nonpregnant p<0.0001) and BVDV (Ag-/Ab-) heifers. No significant differences in average CR between PI BVDV (Ag+/Ab-) heifers and BVDV (Ag-/Ab-) heifers were found. The differences in average FIT between BVDV (Ag+/Ab+; p<0.0001; nonpregnant p<0.0001) and BVDV (Ag-/Ab-) heifers were important statistically. Although there were no BVDV (Ag+/Ab+) pregnant heifers, the differences in average CR between BVDV (Ag+/Ab+) pregnant heifers and BVDV (Ag-/Ab-) heifers were found to be statistically important (p<0.0001). We conclude that fertility is affected in heifers with BVDV (Ag-/Ab+, Ag+/Ab- and Ag+/Ab+).

Relationships among jumping performances and sprint parameters during maximum speed phase in sprinters.

The purpose of this study was to investigate the relationships among jumping performances and speed parameters during maximum speed phase in sprinters. Twenty-one men sprinters volunteered to participate at the beginning of the preparation training phase. All tests-including 100-m sprint running, squat jump (SJ), countermovement jump (CMJ), drop jump (DJ), 60-second repetitive jump (RJ), standing long jump (SLJ), standing triple jump (STJ), standing quintuple jump (SQJ), and standing 10-stride jump (STENJ)-were done on switching mats. Flight (FT) and contact times (CT) during the vertical jump tests and 10-m split times during 100-m sprint running were measured by a 2-channel precision timing system (PTS) connected to the mats. The trace marking method was used for measuring the stride length (SL) through 60 m in 100-m sprint running. Stride frequency (SF), maximum velocity (Vmax), jump height for all vertical jumps, and lower-body power in DJ and RJ were calculated. Statistical analysis showed that the highest significant correlation was found between Vmax and DJ height (r = 0.69; p < 0.05). However, the lowest significant correlation coefficient was found between SL at maximum velocity phase of sprint running and SJ (r = 0.39; p < 0.05). In conclusion, DJ height is demonstrated to be a more effective way to reflect Vmax during sprint running than the other vertical and horizontal jump tests at the beginning of the preparation training phase.

Pathological, serological, and virological findings in sheep infected simultaneously with Bluetongue, Peste-des-petits-ruminants, and Sheeppox viruses.

In this study, pathological, serological and virological examinations were performed on 15 sheep from a flock of 250 sheep and lambs that suffer from simultaneous naturally occurring BTV, PPRV and SPV outbreaks. SPV was diagnosed macroscopically and histopathologically, BTV was diagnosed by ELISA, and PPRV was diagnosed pathologically and by ELISA. Clinically fever, diarrhea, depression, polypnea, conjunctivitis, lacrimation, rhinitis, erosive stomatitis, edema of eyelids, photophobia, cutaneous eruption with erythematous areas especially noticeable in wool-free parts of the body and axilla lesions evolving into papules were observed. At necropsy, the most effected organs were lungs and gut. Subepicardial hemorrhages were also commonly seen. While typical pox lesions were observed in some lambs, usually fibrinous pleuropneumonia was more prominent lung lesion. SPV and PPRV lesions were seen at the histopathological examination of the lesioned tissues, BT lesions were mild than SPV and PPRV microscopically. Serum and leukocyte samples of 15 animals were examined for PPRV and BTV by ELISA; 5 samples were positive for PPRV and 6 BTV, 4 were positive for both PPRV and BTV simultaneously. One hundred animals died, most were lambs. Mortality rates were 100% in lambs and 80% in the herd.

Multispectral co-occurrence with three random variables in dynamic contrast enhanced magnetic resonance imaging of breast cancer.

Presented is a new computer-aided multispectral image processing method which is used in three spatial dimensions and one spectral dimension where the dynamic, contrast enhanced magnetic resonance parameter maps derived from voxel-wise model-fitting represent the spectral dimension. The method is based on co-occurrence analysis using a 3-D window of observation which introduces an automated identification of suspicious lesions. The co-occurrence analysis defines 21 different statistical features, a subset of which were input to a neural network classifier where the assessments of the voxel-wise majority of a group of radiologist readings were used as the gold standard. The voxel-wise true positive fraction (TPF) and false positive fraction (FPF) results of the computer classifier were statistically indistinguishable from the TPF and FPF results of the readers using a one sample paired t-test. In order to observe the generality of the method, two different groups of studies were used with widely different image acquisition specifications.