Explanation and Elaboration Document for the STROBE‐Vet Statement: Strengthening the Reporting of Observational Studies in Epidemiology – Veterinary Extension

The STROBE (Strengthening the Reporting of Observational Studies in Epidemiology) statement was first published in 2007 and again in 2014. The purpose of the original STROBE was to provide guidance for authors, reviewers and editors to improve the comprehensiveness of reporting; however, STROBE has a unique focus on observational studies. Although much of the guidance provided by the original STROBE document is directly applicable, it was deemed useful to map those statements to veterinary concepts, provide veterinary examples and highlight unique aspects of reporting in veterinary observational studies. Here, we present the examples and explanations for the checklist items included in the STROBE‐Vet Statement. Thus, this is a companion document to the STROBE‐Vet Statement Methods and process document, which describes the checklist and how it was developed.     

Interaction of Crop Load and Irrigation on Yield,Fruit Size,Color and Stem-end Splitting Ratio of Apple c.v. ‘Gala,Galaxy’

In this study, the interaction between crop load and irrigation level on yield, fruit size, skin color and stem-end splitting fruit ratio in the apple cultivar ‘Gala, Galaxy’ grafted on rootstock M9 were investigated. Six irrigation programs were applied during the whole growth season: deficit irrigation (rates of 0.25 kc, 0.50 kc, 0.75 kc), full irrigation (rate of 1.00 kc), excess irrigation (rate of 1.25 kc) and non-irrigation (rates of 0.00 kc of “Class A” pan evaporation coefficient). Four crop loads in each irrigation application were performed by hand thinning after the June drop as a- a low crop load (3 fruits cm^?2 TCA), b- a medium crop load (5 fruits cm^?2 TCA), c- a heavy crop load (7 fruits cm^?2 TCA), and d- an un-thinned crop load (>?7 fruits cm^?2 TCA). The total tree yield increased with crop load and irrigation levels. Fruit size was significantly increased by the low crop load. Irrigation increased the fruit size compared to non-irrigation treatment. Further 0.75 kc, 1.00 kc and 1.25 kc irrigation treatments significantly increased the fruit length. Irrigation reduced the fruit flesh firmness. While the low crop load increased the skin red color, it decreased the fruit skin brightness. The yellowness of skin decreased with increasing in the irrigation amount. Irrigation reduced the skin brightness and yellowness, but it increased red color. Crop load and irrigation significantly affected the stem-end splitting fruit ratio. While the splitting fruit ratio increased with a decrease in the crop load, it decreased with an increase in irrigation amount, relatively. Consequently, the low and medium crop load treatments would be beneficial to increase the ratio of marketable fruits without any significant losses in yield for ‘Gala’ apple, especially under 0.75 kc deficit irrigation treatment.     

Antiapoptotic activity of bovine herpesvirus type-1 (BHV-1) UL14 protein

Viruses have evolved different strategies to interfere with apoptotic pathways in order to halt cellular responses to infection. One previous study showed that transient transfection of bovine herpesvirus type-1 (BHV-1) UL14 protein is efficient in protecting Madin Darby kidney (MDBK) and human chronic myelogenous leukemia (K562) cells from sorbitol-induced apoptosis. This protein corresponds to a putative protein of BHV-1, which shares aminoacid sequence with a part of the peptide-binding domain conserved in human heat shock protein (HSP70) family. The pBK-CMV-UL14 plasmid transfected MDBK cells treated with sorbitol did not show caspase-3 and caspase-9 activation with respect to non-transfected MDBK cells (UL14 negative). Furthermore, we report that the expression of the full length sequence of BHV-1 UL14 is evident after 7 h of infection of BHV-1 on MDBK cells which were then treated with sorbitol. These results indicate that UL14 gene product has important implications to enhance cell survival in response to apoptotic stimuli.     

Importance of the rearing period for laying hens in alternative systems

Feather pecking and cannibalism are still major problems in alternative systems for laying hens. Literature and practical experience indicate that unfavourable rearing conditions might be important risk factors for the occurrence of these behavioural disturbances during the laying period.Typical rearing conditions of laying hens from 50 rearing units in Germany and Austria are presented. Obvious risk factors during rearing for feather pecking and cannibalism during the laying period were found. Most flocks were kept under high stocking density (mean: 15 pullets per m' useable area) and some flocks had access to litter only after the second week of life or access to raised perches after the fourth week of life. Plumage condition of pullets and laying hens varied widely in non-beak-trimmed as well as in beak-trimmed flocks.The percentage of pullets with damaged plumage was higher in beak-trimmed than in non-beak-trimmed flocks (medians: 53 % versus 30 %, p = 0,022). In laying hens there was a higher percentage of hens with plumage damage in non-beak-trimmed flocks compared to beak-trimmed flocks (medians: 23 % versus 50 %, p = 0,007). Data analysis will be continued, especially with regard to particular risk factors.     

The effect of veterinary-treated clinical mastitis and pregnancy status on culling in Swedish dairy cows

The interaction of the effects of pregnancy status and veterinary-treated clinical mastitis on culling in Swedish dairy cattle was analyzed with survival analysis. The data were from 978,780 cows with first calvings between 1988 and 1996. Four breeds (Swedish Red and White (SRB), Swedish Friesian (SLB), Swedish Polled Breed and Jersey) were included in the analysis, together with the SRB x SLB crossbreds. Length of productive life was defined as the number of days between first calving and culling or censoring (end of data collection). The model (Weibull proportional hazard) included the interaction of parity by pregnancy status by veterinary-treated clinical mastitis, peak test-day milk-yield deviation within herd-year-parity, age at first calving, year by season, region, breed, herd production level, and the random effect of herd. The effects of pregnancy status and veterinary-treated clinical mastitis were modeled as time-dependent covariates. The lactation was divided into five stages during which a veterinary-treated clinical mastitis and culling might occur and in which the pregnancy status was assumed to be known and culling could occur. Open cows had a pronounced effect on culling: they had a very high risk of being culled in all lactations, and it was even higher if they were treated for mastitis in early lactation. For pregnant cows, the later they got pregnant during the lactation, the greater their risk to be culled. The risk associated with cases of veterinary-treated clinical mastitis remained important throughout the lactation.     

Experimental infection of C3H/HeJ mice with the NY18 isolate of Anaplasma phagocytophilum

Human granulocytic anaplasmosis (HGA), an emerging disease of public health concern in many areas of the world, is caused by Anaplasma phagocytophilum. Small animal models of A phagocytophilum in laboratory mice have been developed and used to study the pathogenesis of HGA. In this study, we characterized the pathologic changes in acute infection of C3H/HeJ mice experimentally infected with the NY18 isolate of A phagocytophilum. Although no clinical signs were noted, acute infection was associated with gross splenomegaly, microscopic inflammatory lesions in the lung and liver, hyperplastic lesions on the spleen, and clinical pathology abnormalities including neutropenia and monocytosis. This study emphasizes the use of well-defined animal models as a valuable tool for the study of A phagocytophilum infections.     

Intra-tumoral gene delivery of feIL-2, feIFN-gamma and feGM-CSF using magnetofection as a neoadjuvant treatment option for feline fibrosarcomas: a phase-I study

Despite aggressive pre- or postoperative treatment, feline fibrosarcomas have a high relapse rate. In this study, a new treatment option based on immune stimulation by intra-tumoral delivery of three feline cytokine genes was performed. The objective of this phase-I dose-escalation study was to determine a safe dose for further evaluation in a subsequent phase-II trial. Twenty-five client-owned cats with clinical diagnosis of fibrosarcoma - primary tumours as well as recurrences - entered the study. Four increasing doses of plasmids coding for feIL-2, feIFN-gamma or feGM-CSF, respectively, were previously defined. In groups I, II, III and IV these doses were 15, 50, 150 and 450 microg per plasmid and a corresponding amount of magnetic nanoparticles. Two preoperative intra-tumoral injections of the magnetic DNA solution were followed by magnetofection. A group of four control cats received only surgical treatment. Side effects were registered and graded according to the VCOG-CTCAE scale and correlated to treatment. Statistical analyses included one-way anova, post hoc and Kruskal-Wallis tests. ELISA tests detecting plasma feIFN-gamma and plasma feGM-CSF were performed. One cat out of group IV (450 microg per plasmid) showed adverse events probably related to gene delivery. As these side effects were self-limiting and occurred only in one of eight cats in group IV, this dose was determined to be well tolerable. Altogether six cats developed local recurrences during a 1-year observation period. Four of these cats had been treated with dose IV. Regarding these observations, a subsequent phase-II trial including a representative amount of cats should be tested for the efficacy of dose IV as well as dose III.     

Blood vitamin concentrations in privately owned dogs fed non-standardized commercial diets and after intake of diets with specified vitamin concentrations

The objective was to investigate in a survey study the blood vitamin concentrations in healthy dogs fed non-specified commercial complete diets and in an intervention study to determine the effects of defined dietary vitamin intakes on blood vitamin levels and hair and skin condition. Sixty-four privately owned dogs, aged from 1 to 8 years, without history of skin or coat problems were included. All animals were fed commercial complete diets with uncertain vitamin concentrations before enrolment. The animals were assigned, according to weight and gender, to four groups with graded vitamin intakes. The blood vitamin levels and skin and coat quality of the dogs were investigated at days 0 and day 122. Coat and hair condition was not influenced by the experimental diets. The retinol concentrations were reduced at the end of the experiment compared with the baseline levels, retinyl esters were not influenced. 25-Hydroxycholecalciferol decreased in all groups, alpha-tocopherol was constant or tended to decrease. Ascorbic acid, thiamine pyrophosphate and riboflavin concentrations were not affected by treatment, flavin adenine dinucleotide and pyridoxal-5'-phosphate were partially reduced on day 122. Cobalamin, pantothenate and biotin concentrations increased with higher dietary intakes, folate levels in tendency. In conclusion, this study gives a survey of blood vitamin concentrations in healthy dogs and provides a data base for the evaluation of the vitamin status in health and disease.