ABSTRACT
African Swine Fever (ASF) has been spreading in numerous southeast Asian countries since a major incursion in mainland China in 2018. Timor-Leste confirmed an outbreak of ASF in September 2019 which resulted in high mortalities in affected pigs. Pigs in Timor-Leste are the second most common type of livestock kept by villagers and represent a traditionally important source of income and prestige for householders. In order to understand the extent of ASF infected villages in Timor-Leste a prevalence survey was designed and conducted in November-December 2019. Timor-Leste has limited laboratory facilities and access to qPCR diagnostic tests. Therefore, a loop mediated isothermal amplification (LAMP) assay was used to detect ASF positive blood samples collected during the prevalence survey. The LAMP assay was proven to be a robust, highly specific and sensitive laboratory test for ASF suitable for use in the field and where there are limited laboratory facilities. The results of the prevalence survey allowed the extent of the ASF incursion to be delineated and the introduction of a disease response strategy to limit the spread of ASF and assist in the recovery of the pig population in Timor-Leste.
ABSTRACT
Recent outbreaks of African swine fever virus (ASFV) have seen the movement of this virus into multiple new regions with devastating impact. Many of these outbreaks are occurring in remote, or resource-limited areas, that do not have access to molecular laboratories. Loop-mediated isothermal amplification (LAMP) is a rapid point of care test that can overcome a range of inhibitors. We outline further development of a real-time ASFV LAMP, including field verification during an outbreak in Timor-Leste. To increase field applicability, the extraction step was removed and an internal amplification control (IAC) was implemented. Assay performance was assessed in six different sample matrices and verified for a range of clinical samples. A LAMP detection limit of 400 copies/rxn was determined based on synthetic positive control spikes. A colourmetric LAMP assay was also assessed on serum samples. Comparison of the LAMP assay to a quantitative polymerase chain reaction (qPCR) was performed on clinical ASFV samples, using both serum and oral/rectal swabs, with a substantial level of agreement observed. The further verification of the ASFV LAMP assay, removal of extraction step, implementation of an IAC and the assessment of a range of sample matrix, further support the use of this assay for rapid in-field detection of ASFV.
Subject(s)
African Swine Fever Virus/genetics , African Swine Fever/epidemiology , African Swine Fever/virology , Disease Outbreaks , Molecular Diagnostic Techniques , Nucleic Acid Amplification Techniques , African Swine Fever/diagnosis , African Swine Fever Virus/isolation & purification , Animals , Female , Male , Molecular Diagnostic Techniques/methods , Molecular Diagnostic Techniques/standards , Nucleic Acid Amplification Techniques/methods , Nucleic Acid Amplification Techniques/standards , Real-Time Polymerase Chain Reaction , Sensitivity and Specificity , Swine , ViremiaABSTRACT
Reports of cystic thyroid and parathyroid masses in cats are uncommon. Herein, the authors describe a series of four cats with cystic ventral cervical lesions, among them thyroid cyst (n=1), thyroid cystadenoma (n=2), and parathyroid adenocarcinoma (n=1). Presentations ranged from completely asymptomatic cervical swellings to signs related to local compression of adjacent structures (e.g., trachea). Ultrasonographic evaluation was helpful in localization of the mass in two cases. Hormone analysis and concentration of cystic fluid were performed in one cat. Surgical excision was performed successfully in three cases. Histopathological examination was performed in all four cases. Long-term prognosis was excellent for those cases in which follow-up was available.
Subject(s)
Adenocarcinoma/veterinary , Cat Diseases/diagnosis , Cysts/veterinary , Parathyroid Neoplasms/veterinary , Thyroid Diseases/veterinary , Adenocarcinoma/diagnosis , Animals , Cat Diseases/diagnostic imaging , Cat Diseases/pathology , Cat Diseases/surgery , Cats , Cystadenoma/diagnosis , Cystadenoma/veterinary , Cysts/diagnosis , Diagnosis, Differential , Female , Male , Parathyroid Neoplasms/diagnosis , Radiography , Thyroid Diseases/diagnosis , Thyroid Neoplasms/diagnosis , Thyroid Neoplasms/veterinary , UltrasonographyABSTRACT
Two Great Danes were evaluated for signs of congestive heart failure. Dilated cardiomyopathy was diagnosed in each dog. Both dogs were diagnosed concurrently with primary hypothyroidism. Following appropriate supplementation with levothyroxine, both dogs showed significant improvement in myocardial contractility, as demonstrated by increased fractional shortening and reductions in left atrial size, and left ventricular end-systolic and diastolic diameters. These observations suggest that hypothyroidism may lead to myocardial failure in Great Danes that is partially or fully reversible following thyroid hormone replacement therapy, and that hypothyroidism should be considered in all Great Danes presenting with systolic failure.
