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This manuscript presents a protocol for a meta-epidemiological study on the characteristics of scientific articles on COVID-19 published during the initial three months of the pandemic. TITLE Characteristics of scientific articles on COVID-19 published during the initial three months of the pandemic: protocol for a meta-epidemiological study AUTHORS Nicola Di Girolamo and Reint Meursinge Reynders ORCID NUMBERS Nicola Di Girolamo ORCID: 0000-0001-5203-9765 Reint Meursinge Reynders ORCID: 0000-0002-2233-9748 AFFILIATIONS AUTHORS Nicola Di Girolamo 1,2 (Associate professor and editor) 1 Center for veterinary health sciences, Oklahoma State University, 2065 W. Farm Road, Stillwater, Oklahoma 74078, USA. 2 EBMVet, Via Sigismondo Trecchi 20, Cremona CR 26100, Italy. Email: nicoladiggi@gmail.com Reint Meursinge Reynders 3,4 (Senior scientist and associate editor) 3 Department of oral and maxillofacial surgery, Amsterdam University Medical Center, University of Amsterdam, Meibergdreef 9, 1105 AZ Amsterdam, The Netherlands. 4 Private practice of orthodontics, Via Matteo Bandello 15, 20123 Milan, Italy. Email: reyndersmail@gmail.com CORRESPONDING AUTHOR Nicola Di Girolamo1,2 (Associate professor and editor) 1 Center for veterinary health sciences, Oklahoma State University, 2065 W. Farm Road, Stillwater, Oklahoma 74078, USA. 2 EBMVet, Via Sigismondo Trecchi 20, Cremona CR 26100, Italy. Email: nicoladiggi@gmail.com ABSTRACT Objectives: (1) What is the prevalence of primary and secondary articles on COVID-19 in the first 3 months of the pandemic? (2) What are the characteristics of both primary and secondary articles on COVID-19? (3) How does the COVID-19 publishing pattern compare with the publishing pattern of the last pandemic, i.e., Swine Flu pandemic? Design: Meta-epidemiological cross-sectional study Study sample: All research articles on Covid-19 indexed in PubMed (Medline) up to 2 April 2020. Main outcome measures: Predominantly prevalence statistics KEYWORDS Covid-19; Coronavirus; SARS-nCoV-2; Swine Flu; H1N1 strain; study design; research quality; healthcare policy; evidence-based medicine INTRODUCTION The outbreak of the coronavirus disease 2019 (COVID-19) pandemic has led to prolific publishing on this health issue. Here, we will assess the scientific literature to analyze the characteristics of articles published on COVID-19 during the first 3 months of this pandemic. We will assess the type, quantity, and quality of this literature to get an insight in the available knowledge base for controlling this pandemic early on. We will divide the literature in primary and secondary articles. Primary articles refer to original research studies that create new or confirming data on a problem, e.g. an health issue. Secondary articles use these data to assess the current knowledge status on this problem. We will also compare the publishing pattern during the first 3 months of the COVID-19 pandemic with this pattern during the most recent pandemic, i.e., the H1N1 pandemic. OBJECTIVES Our main objectives are summarized in the following 3 research questions: (1) What is the prevalence of primary and secondary articles on COVID-19 in the first 3 months of the pandemic? (2) What are the characteristics of both primary and secondary articles? (3) How does the publishing pattern during the first 3 months of the COVID-19 pandemic compare with the publishing pattern of the last pandemic, i.e., Swine Flu pandemic? METHODS Type of study We will conduct a cross-sectional (meta-epidemiological) study. Reporting of the study We will use Strengthening the Reporting of Observational Studies in Epidemiology (STROBE) Statement as the framework for reporting this study (von Elm 2007). We will report our methods for the Swine Flu sample at the end of the methods section. Eligibility criteria Any type of article published on COVID-19 will be eligible. This implies that a broad spectrum of articles ranging from letters to the editors to randomized controlled trials will be included. No eligibility criteria will be applied to specific participants, interventions, comparators, outcomes, endpoints, or settings of the articles. Articles that did not present an abstract as well as abstracts that were not reported in English will be excluded. Information sources and search strategy We will search eligible articles on COVID-19 in PubMed (MEDLINE) till April 2 2020. Our search strategy for PubMed (MEDLINE) is presented in Table 1 and will be prepared for the eligible search dates. We will use the search string “(COVID-19 OR COVID)” and apply the filter ‘Abstract’. Table 1. Search strategy for COVID-19 Search: COVID-19 OR COVID Filters: Abstract ((((((("covid 19"[All Fields] OR "covid 2019"[All Fields]) OR "severe acute respiratory syndrome coronavirus 2"[Supplementary Concept]) OR "severe acute respiratory syndrome coronavirus 2"[All Fields]) OR "2019 ncov"[All Fields]) OR "sars cov 2"[All Fields]) OR "2019ncov"[All Fields]) OR (("wuhan"[All Fields] AND ("coronavirus"[MeSH Terms] OR "coronavirus"[All Fields])) AND (2019/12/1:2019/12/31[Date - Publication] OR 2020/1/1:2020/12/31[Date - Publication]))) OR "COVID"[All Fields] Translations COVID-19: "COVID-19"[All Fields] OR "COVID-2019"[All Fields] OR "severe acute respiratory syndrome coronavirus 2"[Supplementary Concept] OR "severe acute respiratory syndrome coronavirus 2"[All Fields] OR "2019-nCoV"[All Fields] OR "SARS-CoV-2"[All Fields] OR "2019nCoV"[All Fields] OR (("Wuhan"[All Fields] AND ("coronavirus"[MeSH Terms] OR "coronavirus"[All Fields])) AND (2019/12[PDAT] OR 2020[PDAT])) Selection of articles and data extraction Two operators (Nicola di Girolamo and Reint Meursinge Reynders) will conduct the selection of articles and the data extraction procedures independently. These operators pilot tested these methods on 40 articles to calibrate operators and to fine-tune research methods, e.g., the data extraction forms (Table 2). Disagreements during the selection of articles and data extraction procedures will be resolved through discussions between these operators. A methodologist will be consulted in the case of persisting disagreements. We will report the flow of our selection of articles and will include our raw data with the final research study. Table 2. Data extraction forms Title: Report the title of the article PMID: Report the PubMed-Indexed for MEDLINE (PMID) Authors: Report the authors of the article Citation: Report the citations of the article First author: Report the first author of the article Journal/Book: Report the name of the journal or book Publication year: Report the year of publication of the article Creation date: Report the date of the creation of the Digital Object Identifier (DOI) DOI: Report the DOI DOI LINK: Report the link to the DOI Language of full text: Report the language in which the full text of the article was published Language if not English: Report the language of the full text if not English Country: Report the country of the first institution of the first author Type of study: Report the type of study, e.g., human medical research/in vitro/in silico/review/guidelines etc. Study design: Report the study design. Only the design of human medical research studies will be reported Sample size: Report the sample size. Only the sample size of human medical research studies will be reported Objectives: Report whether the objectives of the article were reported in the abstract. Possible answers: Yes/No Limitations/problems reported in the abstract: Report whether limitations/or problems of the article were reported in the abstract. Possible answers: Yes/No Additional comments: Report possible additional comments Conclusions: Copy and paste the conclusions reported in the abstract Classification of the included articles We will classify each included article as either a primary or a secondary article. Primary articles cover original research studies and secondary articles cover perspectives and syntheses of the available knowledge on COVID-19 such as, viewpoints, commentaries, guidelines, reviews etc. Primary articles will be divided in 5 groups, i.e., human medical research, in silico, in vitro, animal research, and surveys or studies on health professionals. These groups will be further subdivided according to the pertinent research designs. Our classification of articles will not be exclusively based on the labels assigned by the authors of the articles, because of the potential risk of mislabeling (Esene 2014). We will weigh the author’ labels before assigning our final classification. Abstract assessment We will screen the abstracts to assess whether objectives and limitations of the article were reported or not. Limitations will be subdivided in methodological and general limitations. The former refers to reporting of at least one methodological limitation in the abstract. General limitations refer to reporting some sort of limitation not inherent to the design of the paper, e.g., further research on this topic is needed etc. Information sources and search strategy for the Swine Flu sample We will search PubMed (MEDLINE) for eligible articles on H1N1 during the first 3 months of the Swine Flu outbreak (04/25/09-7/25/09). Our search strategy for PubMed (MEDLINE) is presented in Table 3. We will use the search string “((H1N1) OR "swine influenza")” and apply the filter ‘Abstract’. This search strategy will be prepared for the eligible search dates. Table 3. Search strategy for H1N1 Search: (H1N1) OR (swine influenza) Filters: Abstract "H1N1"[All Fields] OR (((("orthomyxoviridae infections"[MeSH Terms] OR ("orthomyxoviridae"[All Fields] AND "infections"[All Fields])) OR "orthomyxoviridae infections"[All Fields]) OR ("swine"[All Fields] AND "influenza"[All Fields])) OR "swine influenza"[All Fields]) Translations swine influenza: "orthomyxoviridae infections"[MeSH Terms] OR ("orthomyxoviridae"[All Fields] AND "infections"[All Fields]) OR "orthomyxoviridae infections"[All Fields] OR ("swine"[All Fields] AND "influenza"[All Fields]) OR "swine influenza"[All Fields] Selection of articles, data extraction, and classification of the included articles for the Swine Flu sample For the selection of articles, data extraction, and the classification of the included articles for the Swine Flu sample we will adopt the methods presented for the COVID-19 sample. We will report on the classification of primary and secondary articles in this sample. Data analysis We will use descriptive statistics expressed as medians with interquartile ranges (IQR) and ranges or absolute counts and percentages. Multivariate logistic regression models will be developed to explore factors associated with the primary outcomes and to provide odds ratio adjusted for confounders (Peng & So, 2002; Heck, Thomas & Tabata, 2013). Variables will be retained in the models regardless of their statistical significance. Goodness of fit will be assessed with Hosmer-Lemshow test and Nagelkerke R squared. Data analyses and figures will be done with SPSS (version 24, IBM) and R 3.6.3 (R Core Team, 2020, www.R-project.org/). Outcomes Table 4 summarizes our planned outcomes. For most of these outcomes we will calculate prevalence statistics. We did not involve patients or other stakeholders in developing our research questions or outcomes. Table 4. Summary of planned outcomes Published papers on COVID-19 Primary and secondary articles on COVID-19 Type of primary articles on COVID-19 Research design of primary articles on COVID-19 Type of secondary articles on COVID-19 Articles on COVID-19 per country Journals on COVID-19 Reporting of objectives in the abstract in included articles on COVID-19 Reporting of limitations in the abstract in included articles on COVID-19 Published papers on H1N1 Primary and secondary articles on H1N1 Type of primary articles on H1N1 Research design of primary articles on H1N1 Type of secondary articles on H1N1 Differences in publishing trends between COVID-19 and H1N1 Differences between the protocol and the final study Originally we also planned to extract whether included studies were part of multi center projects. After the completion of our pilot tests we decided not to extract these data, because they could not be extracted reliably. This manuscript represents the latest version of our protocol, i.e., the protocol after the completion of our pilot tests. All differences between this protocol and the methods used in the study will be reported in the final manuscript of the study with rationale. REFERENCES Esene 2014 Esene IN, Ngu J, El Zoghby M, et al. Case series and descriptive cohort studies in neurosurgery: the confusion and solution. Childs Nerv Syst. 2014;30(8):1321–1332. doi:10.1007/s00381-014-2460-1. Von Elm 2007 von Elm E, Altman DG, Egger M, Pocock SJ, Gøtzsche PC, Vandenbroucke JP; STROBE Initiative. The Strengthening the Reporting of Observational Studies in Epidemiology (STROBE) statement: guidelines for reporting observational studies. Ann Intern Med. 2007 Oct 16;147(8):573-7.

The purpose of the present meta-epidemiological study is (1) to identify the proportion of authors that, based on their self-declared contributions, did not met the minimum criteria for authorship as established by the ICMJE and McNutt recommendations, (2) to quantify the difference between these two recommendations, and (3) to identify the proportion of authors that contributed exclusively funding or materials to a study. For this reason, we will perform a cross-sectional study of CRediT statements published in scholarly articles. We will contact three publishing companies, the Public Library of Sciences (PLoS), Elsevier, and Wiley to obtain CRediT statements. We will ask the publishers to provide every CRediT statement for every author of any article published in any of their journals. We will classify whether authors fulfilled minimum authorship recommendations advised by the ICMJE and by McNutt et al (2008) depending on the combination of items that they declared to have contributed in the CRediT statements. In addition, we will classify whether CRediT statements were suggestive of one specific authorship malpractice, that we defined “supply authorship”, and is characterized by providing exclusively funding or materials to a scholarly article. In order to define the combinations of items that are consider the minimum contribution to fulfil authorship recommendations, two investigators (ND, RMR) will independently evaluate the criteria for authorship (ICMJE, McNutt, “supply”) and develop a logical operation of CRediT items (using AND and OR) for each criterion. If there is persistent disagreement regarding the combinations, a third operator with methodological experience on authorship (VL) will be consulted as an arbiter. The primary outcomes of this study will be: - The proportion of authors that provided CRediT statements that did not suffice to justify authorship based on ICMJE recommendations - The proportion of authors that provided CRediT statements that did not suffice to justify authorship based on McNutt recommendations - The difference in these proportions - The proportion of authors that based on the CRediT statements provided only funding and materials (supply authors)

Additional file 2. Data collection forms, quality checklists, and summary of findings tables.

Additional file 1. Search strategy.

OBJECTIVE To explore sources of serum aldosterone concentration variability in a population of healthy and diseased ferrets, determine a preliminary 1 -sided reference interval for serum aldosterone concentration in healthy ferrets, and identify a decision limit to differentiate healthy from diseased ferrets on the basis of serum aldosterone concentration. DESIGN Prospective threshold definition and diagnostic accuracy study. ANIMALS 78 healthy (n = 56) and diseased (22) ferrets. PROCEDURES Serum aldosterone concentrations were measured on consecutively admitted ferrets, and an upper reference limit for aldosterone concentrations was established. Sensitivity and specificity of aldosterone concentration cutoffs to differentiate healthy from diseased ferrets were estimated with receiver operating characteristic curve analysis. RESULTS Measurements of serum aldosterone concentrations in the ferrets showed wide variability, with a median concentration of 4.75 pg/mL (interquartile range, 0.55 to 17.9 pg/mL; range, 0.02 to 283.9 pg/mL) and 76% (59/78) of samples having concentrations < 18 pg/mL. Ferrets that were healthy, older, or sexually inactive had significantly lower aldosterone concentrations. The upper limit of the reference interval for healthy ferrets was 13.3 pg/mL (90% confidence interval, 9.9 to 16.9 pg/mL). Analysis of receiver operating characteristic curves indicated that an aldosterone concentration cutoff value of 7.6 pg/mL differentiated healthy ferrets from diseased ferrets with a sensitivity of 72.7% and specificity of 73.2% (area under the curve, 0.79; 95% confidence interval, 0.67 to 0.91). CONCLUSIONS AND CLINICAL RELEVANCE Results suggested that high aldosterone concentrations should not be considered diagnostic of primary hyperaldosteronism in ferrets. A need exists to develop better tests to identify primary hyperaldosteronism.

Background Titles and abstracts are the most read sections of biomedical papers. It is therefore important that abstracts transparently report both the beneficial and adverse effects of health care interventions and do not mislead the reader. Misleading reporting, interpretation, or extrapolation of study results is called “spin”. In this study, we will assess whether adverse effects of orthodontic interventions were reported or considered in the abstracts of both Cochrane and non-Cochrane reviews and whether spin was identified and what type of spin. Methods Eligibility criteria were defined for the type of study designs, participants, interventions, outcomes, and settings. We will include systematic reviews of clinical orthodontic interventions published in the five leading orthodontic journals and in the Cochrane Database. Empty reviews will be excluded. We will manually search eligible reviews published between 1 August 2009 and 31 July 2019. Data collection forms were developed a priori. All study selection and data extraction procedures will be conducted by two reviewers independently. Our main outcomes will be the prevalence of reported or considered adverse effects of orthodontic interventions in the abstract of systematic reviews and the prevalence of “spin” related to these adverse effects. We will also record the prevalence of three subtypes of spin, i.e., misleading reporting, misleading interpretation, and misleading extrapolation-related spin. All statistics will be calculated for the following groups: (1) all journals individually, (2) all journals together, and (3) the five leading orthodontic journals and the Cochrane Database of Systematic Reviews separately. Generalized linear models will be developed to compare the various groups. Discussion We expect that our results will raise the awareness of the importance of reporting and considering of adverse effects and the presence of the phenomenon of spin related to these effects in abstracts of systematic reviews of orthodontic interventions. This is important, because an incomplete and inadequate reporting, interpretation, or extrapolation of findings on adverse effects in abstracts of systematic reviews can mislead readers and could lead to inadequate clinical practice. Our findings could result in policy implications for making judgments about the acceptance for publication of systematic reviews of orthodontic interventions.

An acceptable agreement permits interchangeability of the instruments. For this purpose, we have investigated the agreement of several clinical instruments frequently used in clinical practice with their laboratory counterpart. We have estimated the agreement between a point-of-care blood gas analyzer (i-Stat, Abaxis) and a bench-top blood gas analyzer (Nova, Biomedical) in venous samples from Hermann’s tortoises. We have estimated the agreement between a point-of-care chemistry analyzer (VetScan VS2, Abaxis) and a laboratory analyzer (Olympus AU400, Olympus Co.) in venous samples from Hermann’s tortoises. We have estimated the agreement between portable blood glucose meters (Accu-Chek, Aviva; AlphaTrak 2, Abbott) and a laboratory analyzer (Dimension EXL, Siemens) in venous samples from client-owned rabbits. We have estimated the agreement between point-of-care bench-top glucose measurement (VetScan VS2, Abaxis) and a laboratory analyzer (Dimension EXL, Siemens) in venous samples from client-owned rabbits. Beyond method comparison and validation, reference interval determination for common laboratory testing is required to allow the clinician to discriminate individuals that are different from the remaining population for a certain parameter. We have calculated reference intervals for blood gas in Hermann’s tortoises. We have calculated reference intervals for protein electrophoresis in Hermann’s tortoises. We have described normal hematology in Hermann’s tortoises. We have calculated reference intervals for clinical chemistry in Hermann’s tortoises. We have calculated reference intervals for aldosterone in ferrets. Based on our results, animal species requires individual validation of laboratory methods and reference intervals. Lack of consideration of these findings may result in clinical misdiagnosis and improper treatment of animals.

In 2011, several outbreaks of West Nile disease occurred in Sardinia (Italy). The region hosts several chelonian species. Because of the increasing concern on the potential role that ectotherms may play in the ecology of West Nile virus (WNV), in October 2011 blood samples were collected from 41 endemic Sardinian chelonians and tested for the presence of active WNV infection or neutralizing antibodies by real time polymerase chain reaction (RT-PCR) and serumneutralisation, respectively. Neither WNV neutralising antibodies (0%; 95% CI: 0‑8.4%) nor WNV RNA (0%; 95% CI: 0‑6.8%) were found in the tested samples. According to the results of this screening survey, it is unlikely that chelonians are involved in the epidemiology of the 2011 WNV outbreaks in Sardinia.

Antibiotic therapy represents an essential aid in the treatment of sick or injured loggerhead sea turtle (Caretta caretta,). Marbofloxacin is commonly used in loggerhead sea turtles at 2 mgjkg similarly to other species. The aim of the study was to increase our understanding of marbofloxacin pharmacokinetic in Caretta caretta. Six healthy turtles were divided in two groups: the IV group (subjects A, B, D) and the IM group (subjects C, H, F). Each animal received a single dose of MarbocyP 2%, corresponding to 2 mg/ kg of marbofloxacin. The drug was administered via intravenous (IV) and intramuscular (IM) routes. After the administration, 13 blood samples were collected at different times:for IVgroup at 0, 10, 20, 30, 45 minutes, 1, 2, 4, 8, 12 hours and 1, 2, 2,5 days; for IM group at 0, 30 minutes, 1, 1.5, 2, 3, 4, 6, 8, 12 hours, and 1, 2, 2,5 days. A HPLC-UV (k 295 nm) analytical method was used to measure marbofloxacin concentrations in blood. The pharmacokinetic behavior of the drug was best described by a non-compartmental model The plasma concentration profiles of marbofloxacin were similar for the two administration routes and the residual concentrations at 1 day were almost identical. The molecule was detected up to 2,5 days. The apparent volume of distribution was > 1 L/kg. The IM route resulted in a rapid and complete marbofloxacin absorption (F = 131, 43 ±12,06%).

Data on proteinuria are lacking in giraffes, therefore the aim of our research was to study urine proteome with sensitive analytical methods to obtain important information regarding not only the kidney function but also the general health status. Twenty-nine samples were collected during 2013 and 2014, through non-invasive methods (from the ground by a syringe) from 4 specimens of Giraffa camelopardalis. Urinary total proteins (UTP), creatinine and the urine protein to creatinine ratio (UP/C) were measured by automated methods (Olympus AU400) routinely used in small animal practice. Urinary proteins were separated by sodium-dodecyl-sulphate polyacrilamide gel electrophoresis (SDS-PAGE) and gels were stained with silver nitrate. Mean UTP±SD was 12.1±0.6 mg/dL ranging from 2.4 to 30 mg/dL, while mean UP/C values were 0.06±0.01. SDS-PAGE yielded an overall view of the protein profile allowing the separation of 7±2 protein bands. Electropherograms showed few protein bands as compared to dog and cat and big felids. The most represented band had an apparent molecular weight (MW) of 67-69 kDa and could be identified as putative albumin. Between 65 and 14 kDa only few low abundance bands were present, while more intense bands had a MW lower than 14 kDa. Our data could be considered as representative of animals with no clinical signs of disease resulted in a useful diagnostic tool that could help clinicians in quantitative and qualitative evaluation of proteinuria also in giraffes.

Data on proteinuria in giraffes are lacking, therefore the aim of our research was to study urine proteome with sensitive analytical methods to obtain important information regarding not only the kidney function but also the general health status. Fifty urine samples were collected during 2013 and 2014, through non-invasive methods from 21 specimens of Giraffa camelopardalis. Urinary total proteins (UTP), creatinine and urine protein to creatinine ratio (UP/C) were measured by automated methods routinely used in small animal practice. Urinary proteins were separated by sodium-dodecyl-sulphate polyacrylamide gel electrophoresis (SDS-PAGE) and gels were stained with silver nitrate. Mean UTP±SD was 16.1±8.8 mg/dL (range 2.4-30 mg/dL), while mean UP/C was 0.08±0.03 (range 0.04-0.2); these values can be considered representative of healthy animals. SDS-PAGE yielded an overall view of the urine protein profile allowing the separation of 6±2 protein bands. Electropherograms showed few protein bands as compared to dogs, cats (Ferlizza et al., 2012) and big felids (Di Cerbo et al., 2013). The most represented band had an apparent molecular weight (MW) of 67-69 kDa and could be identified as putative albumin. Between 65 and 14 kDa only few low abundance protein bands were present, while more intense bands had a MW lower than 14 kDa. This is the first study reporting UTP and UPC ranges in giraffes. 1-D-SDS-PAGE yielded an overall view of the protein profile and resulted in a useful diagnostic tool that could help clinicians in qualitative evaluation of proteinuria. References Di Cerbo P., Ferrari G., Isani G., Ferlizza E., Friedrich K.G., 2013. Urine proteome in big cats (Panthera leo, Panthera tigris, Panthera pardus). In: proceedings of the International Conference on Diseases of Zoo and Wild Animals, Vienna, A. Ferlizza E., Armuzzi E., Buono A., Carpenè E., Andreani G., Dondi F., Isani G., 2012. Urine proteome from healthy dogs, cats, horses and cows. In: proceedings of the European Society of Veterinary Clinical Pathology Conference, Ljubljana, SLO.

Asian palm civets (Paradoxurus hermaphroditus), or toddy cats, belong to the family Viverridae. Little is known about the pathology of these animals and few articles have been published, mainly concerning their important role as wild reservoir hosts for severe infectious diseases of domestic animals and human beings. A 4-year-old, female Asian palm civet was found dead by the owner. At necropsy, large amount of adipose tissue was found in the subcutis and in the peritoneal cavity. Most of the pancreas appeared red, translucent. Hepatomegaly, discoloration of the liver were evident, with multifocal areas of degeneration, characterized by white nodular lesions. Histologically, the pancreas showed severe interstitial and perilobular necrosis and extensive haemorrhages, with separation of the interstitium, mild reactive inflammation at the periphery of the pancreatic lobules. Liver showed multifocal foci of vacuolar degeneration, lipidic accumulation, sometimes associated to hepatocyte necrosis. A diagnosis of acute severe hemorrhagic-necrotizing pancreatitis (or acute pancreatic necrosis) associated with pancreatic and hepatic lipidosis was made. To the best of our knowledge, this represents the first case report of acute lethal pancreatitis in an Asian palm civet. Although the exact cause of the disease remains undetermined, a hypothesis of the cause and pathogenesis is discussed, pointing out dietary indiscretion and consequent overweight as possible important risk factors.

Congenital heart diseases have rarely been described in rabbits. The purpose of the present case report is to describe the clinical, radiographic, echocardiographic, and pathological features of a partial atrioventricular septal defect in a pet rabbit. A 3-month-old, 380-g male vaccinated pet rabbit was presented for decreased activity, increased respiratory rate and effort, anorexia, and decreased fecal output of 2 days duration. Total body radiographic images revealed severe cardiomegaly associated with enlarged caudal pulmonary vessels and increased interstitial to alveolar lung pattern. Echocardiographic imaging showed evidence of distended heart chambers, abnormal flow through the atria, and mitral valve regurgitation. The rabbit was treated with furosemide and an angiotensin-converting enzyme inhibitor but rapidly deteriorated and died. Necropsy confirmed the dilation of both ventricles and the presence of a partial atrioventricular septal defect associated with an ostium primum atrial septal defect just over the tricuspid valve and the mitral valve.

Background A novel bivalent vaccine to protect against myxomatosis and rabbit haemorrhagic disease is commercially available for pet rabbits. Hypothesis/Objectives To describe the appearance of cutaneous lesions arising in pet rabbits positive for myxoma virus (MV) by RT-PCR evaluation shortly after vaccination. Animals Four pet rabbits presenting with papular, crusting skin lesions ~10 days after vaccination. Methods Histological evaluation of formalin-fixed skin biopsies obtained from lesional skin (case 1). Real-time polymerase chain reaction (RT-PCR) evaluation of paraffin-embedded tissue from skin biopsies (case 1) and crusts obtained from the lesion surface (cases 2–4) for myxoma virus are reported as cycle threshold (Ct) values. Results Lesions affecting the ear pinna, dorsal aspect of the nose, vulva and/or conjunctiva are reported. Histopathological findings included severe ulcerative, necrotizing dermatitis and intralesional cytoplasmic inclusion bodies in myxoma cells. DNA was amplified from all the paraffin-embedded skin biopsies (Ct = 34–35) and crusts (Ct = 20–24). Conclusions and clinical importance Although a wild virus challenge cannot be definitively excluded, veterinarians and pet-owners should be aware that cutaneous lesions have been observed after vaccination with this novel vaccine in low numbers of rabbits.

A loggerhead sea turtle (Caretta caretta) was found stranded alive along the Adriatic coast close to Ancona, Italy, displaying obtundation, tachypnea, and increased respiratory effort. It died a few hours after admission, and a postmortem examination was immediately performed. Miliary yellowish nodules were evident in the liver, and a lower number in the heart, stomach, and gut wall. Hundreds of whitish nodules were scattered in the lungs, with the majority of the pulmonary parenchyma being replaced by the lesions. Histologically, all nodular lesions consisted of a small central area of necrosis with acid-fast bacilli surrounded by epithelioid cells, macrophages, and lymphocytes. Giant cells were found in the spleen and the liver. Kidneys, lungs, liver, spleen, brain, and skin lesions were inoculated aseptically onto general isolation media and selective isolation media for mycobacteria. The isolate showed a restriction pattern identical to Mycobacterium chelonae by polymerase chain reaction-restriction fragment length polymorphism. To the best of the authors' knowledge, this is the first description of a disseminated infection caused by a potentially pathogenic mycobacteria in a stranded, free-ranging loggerhead sea turtle. Veterinary staff and biologists who handle sea turtles with suspected mycobacterial disease should protect themselves appropriately.