The current reticuloendotheliosis virus (REV) antibody detection kit that uses enzyme-linked immunosorbent assay (ELISA) needs concentrated virus, which is difficult to obtain due to its poor propagation in cells. In addition, this kit detects only chicken antibody but not other species. To overcome these disadvantages, we cloned and expressed REV env gene to develop monoclonal antibodies (mAbs), which we used for antibody detection in ELISA. Three mAbs were prepared from mice. These three mAbs could recognize REVs from ducks and geese by immunodot assay. In addition, the epitopes that the three mAbs recognized were determined by using three different env protein fragments by western blotting. One mAb was used to develop a blocking ELISA (bELISA) coated with expressed env protein to detect anti-REV antibody in chicken serum. This assay had a 98.8% (79/80) agreement with a commercial ELISA kit. Another 146 chicken sera with known neutralization antibodies were used as positive controls to evaluate this bELISA. The sensitivity and specificity this bELISA were 88.9% (40/45) and 94.8% (91/96), respectively. Thus, this bELISA could be used for anti-REV antibody detection in birds.Detección de anticuerpos contra el virus de la reticuloendoteliosis mediante el ensayo de inmunoabsorción con enzimas ligadas en formato de bloqueo con la expresión de la proteína de de la envoltura.El estuche actual para la detección de anticuerpos contra la reticuloendoteliosis (REV) que utiliza un ensayo de inmunoabsorción con enzimas ligadas (ELISA) necesita virus concentrado, que es difícil de obtener debido a su baja propagación en células. Además, este estuche detecta sólo los anticuerpos de pollo, y no de otras especies. Para superar estas desventajas, se clonó y se expresó el gene env del virus de la reticuloendoteliosis para desarrollar anticuerpos monoclonales, que se utilizaron para la detección de anticuerpos por ELISA. Se prepararon tres anticuerpos monoclonales a partir de ratones. Estos tres anticuerpos pudieron reconocer virus de reticuloendoteliosis de patos y gansos por un ensayo inmunodot . Además, se determinaron los epítopos que los tres anticuerpos monoclonares reconocieron mediante el uso de tres diferentes fragmentos de la proteína env por inmunotransferencia Western. Un anticuerpo monoclonal fue usado para desarrollar un ELISA de bloqueo (bELISA) recubierto con una proteína env expresada y se utilizó para detectar anticuerpos anticuerpos contra el virus de la reticuloendoteliosis en el suero de pollos. Este ensayo mostró una concordancia del 98.8% (79/80) con un estuche comercial de ELISA. Otros 146 sueros de pollos reconocidos por contener anticuerpos neutralizantes fueron utilizados como controles positivos para evaluar este método de ELISA de bloqueo. La sensibilidad y la especificidad del método de ELISA de bloqueo fueron del 88.9% (40/45) y del 94.8% (91/96), respectivamente. Por lo tanto, este método de ELISA de bloqueo podría ser utilizado para la detección de anticuerpos contra el virus de la reticuloendoteliosis en aves.
Canine MDR1 gene mutations produce translated P-glycoprotein, an active drug efflux transporter, resulting in dysfunction or over-expression.The 4-base deletion at exon 4 of MDR1 at nucleotide position 230 (nt230[del4]) in exon 4 makes P-glycoprotein lose function, leading to drug accumulation and toxicity.The G allele of the c.-6-180T＞G variation in intron 1 of MDR1 (single nucleotide polymorphism [SNP] 180) causes P-glycoprotein over-expression, making epileptic dogs resistant to phenobarbital treatment.Both of these mutations are reported to be common in collies.This study develops a more efficient method to detect these two mutations simultaneously, and clarifies the genotype association with the side effects of chemotherapy.Genotype distribution in Taiwan was also investigated.An oligonucleotide microarray was successfully developed for the detection of both genotypes and was applied to clinical samples.No 4-base deletion mutant allele was detected in dogs in Taiwan.However, the G allele variation of SNP 180 was spread across all dog breeds, not only in collies.The chemotherapy adverse effect percentages of the SNP 180 T/T, T/G, and G/G genotypes were 16.7%, 6.3%, and 0%, respectively.This study describes an efficient way for MDR1 gene mutation detection, clarifying genotype distribution, and the association with chemotherapy.
Reverse transcription polymerase chain reaction (RT-PCR) was used routinely to detect the avian influenza virus (AIV) nucleoprotein (NP) gene. The purpose of the present study was to compare the correctness of a nested RT-PCR (nRT-PCR), one conventional RT-PCR with its outer primer (oRT-PCR) and the other conventional RT-PCR with its inner primer (iRT-PCR) to detect AIV NP gene. A total of 365 AI-free fecal swabs (73 pools), 7 tracheal swabs and anllantoic fluid from 25 chicken embryos were used to determine the analytic specificities of those tests. Compared with the iRT-PCR, the nRT-PCR was more sensitive for AIV detection. However, the specificities of nRT-PCR, oRT-PCR and iRT-PCR were 48.6% (35/72), 100% (67/67) and 91.3% (84/92), respectively. The amplifying band was sequenced and confirmed to be the AIV NP gene as the positive control. The specificity of this nRT-PCR is too low to be used for the AIV screening test.
An 18-yr-old, captive-born male Bengal tiger (Panthera tigris tigris) presented a 1-yr history of chronic and progressive paraparesis in both hind limbs. Lateral and hind limb radiographs were revealed normal except for severe spondylosis deformans, forming a bony bridge between the last lumbar and the first sacral vertebra and disc mineralization between the second and third lumbar vertebra. Medical therapies were instituted, including corticosteroids, hydroacupuncture and electroacupuncture. Animal training allowed veterinarians to perform acupuncture safely without having to anesthetize the animal. Animal training made intensive treatment possible. Neither corticosteroids nor hydroacupuncture alone provided much clinical improvement. The tiger reacted positively after electroacupuncture was performed. The tiger began showing clinical improvement after three electroacupuncture treatments and could eventually walk on all four limbs at the end of the treatment.
Abstract : The Formosan sika deer (Cervus nippon taiouanus) is an endemic subspecies in Taiwan. The original wilddeer has been extinct since the late 1960s. The largest captive population is located at the Taipei Zoo. Except forinfectious disease outbreaks, no systemic medical research has been reported for this subspecies. This study wasconducted to analyze the medical status of the captive Formosan sika deer population, including the hematologicaland serum chemistry characteristics. To accomplish this, medical records for 34 Formosan sika deer from January2003 to January 2014 were acquired and analyzed. The most common illness and cause of death was trauma, followedby gastrointestinal and respiratory disease, respectively. The hematologic and serum chemical values of healthy adultswere quite different from those of sika deer (Cervus nippon yesoensis). This study provides a closer medicalunderstanding of this subspecies and the results will facilitate its management.Keywords : Formosan sika deer, gastrointestinal disease, respiratory disease, Taiwan, trauma
Novel clade 2.3.4.4 H5 highly pathogenic avian influenza virus (HPAIV) outbreaks have occurred since early 2015 in Taiwan and impacted the island economically, like they have many countries. This research investigates the immunogenicity of two HPAIV-like particles to assess their promise as vaccine candidates.The haemagglutinin (HA) gene derived from clade 2.3.4.4 H5 HPAIV and matrix protein 1 (M1) gene were cloned into the pFastBac Dual baculovirus vector. The resulting recombinant viruses were expressed in Spodoptera frugiperda moth (Sf)21 cells and silkworm pupae to generate Sf21 virus-like particles (VLP) and silkworm pupa VLP. Two-week-old specific pathogen-free chickens were immunised and their humoral and cellular immune responses were analysed.The silkworm pupa VLP had higher haemagglutination competence. Both VLP types elicited haemagglutination inhibition antibodies, anti-HA antibodies, splenic interferon gamma (IFN-γ) and interleukin 4 (IL-4) mRNA expression, and CD4+/CD8+ ratio elevation. However, chickens receiving silkworm pupa VLP exhibited a significantly higher anti-HA antibody titre in ELISA after vaccination. Although Sf21 VLP recipients expressed more IFN-γ and IL-4, the increase in IFN-γ did not significantly raise the CD4+/CD8+ ratio and the increase in IL-4 did not promote anti-HA antibodies.Both VLP systems possess desirable immunogenicity in vivo. However, in respect of immunogenic efficacy and the production cost, pupa VLP may be the superior vaccine candidate against clade 2.3.4.4 H5 HPAIV infection.
The Formosan serow (Capricornis swinhoei) is endemic to Taiwan. The wild population has declined dramatically over the past few decades and the species is listed as a "precious and rare species" protected under law in Taiwan. Disease investigations have been rare except for sporadic observations of wild individuals, and no long-term disease survey has been performed on this species. The objective of this study was to identify and report on the most common diseases in captive Formosan serows and determine the potential causes. Medical records of Formosan serows (n = 62) housed at the Taipei Zoo over a 20-yr period (1991-2011) were collected and analyzed for this study. The most common diseases affected the gastrointestinal system and the skin. Parasitic etiologies accounted for greater than 85% of these diseases, and coinfection was common. Coccidia and lice were the most common endo- and ectoparasites, respectively. High mortality was noted in serows less than 1 yr old associated with parasitism. The results from this study could provide vital information on disease prevention and species management, which may greatly help in rehabilitation of captive and wild populations.
Confined bears are deprived of natural stimulus, such as large roaming domains, active foraging and enriched habitats, making them vulnerable to develop stereotypic behaviors. Motivation drives and neuro-pathological imbalance are the causes. Various stereotypic behavior types have been reported, including oral, head and locomotory related behaviors, and pacing are the most commonly seen. Stereotypic behavior level could be monitored using an observation-reporting system or fecal corticoid. Environmental and feeding enrichment are the proposed solutions; however, habituation could be developed. Parallel intermittent and consecutive environmental enrichment and unpredictable feeding schedule would be beneficial. Medication, especially long-term and low-dosage fluoxetine, was found to be effective in alleviating bear stereotypic behaviors.
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