Suspected clinical toxoplasmosis in a 12-week-old puppy in Singapore

Abstract Background Toxoplasma gondii is traditionally known as a parasite of felids, with possible infection in intermediate hosts such as dogs and humans, and thus a disease of public health significance. Published data on the prevalence of toxoplasmosis in dogs and cats in Singapore is scanty, and this paper documents a suspect clinical case of toxoplasmosis in a free-roaming puppy trapped from an offshore island of Singapore. Case presentation A 12-week-old puppy presented with hindlimb weakness and sarcopenia, with rapidly progressing ascending paralysis and respiratory distress, one week after trapping. Toxoplasmosis was suspected after indirect fluorescence antibody testing (IFAT) revealed anti-T. gondii antibodies. The puppy responded quickly to clindamycin treatment and was discharged from hospital after 10 days. Conclusion While rare and undocumented, veterinary clinicians in Singapore are advised to also include toxoplasmosis infection as a differential diagnosis in dogs presenting with similar clinical signs. This is especially so for dogs which have access to the outdoors

Robotic left hepatectomy with revision of hepaticojejunostomy

Laparoscopic hepatectomy and hepaticojejunostomy remain a surgical challenge despite the recent advances in minimally invasive surgery. A robotic surgical system has been developed to overcome the inherent limitations of the traditional laparoscopic approach. However, techniques of robotic hepatectomy have not been well described, and a description of robotic major hepatectomy with bilioenteric anastomosis can be found only in two previous reports. Here, we report a 33-year-old man with a history of choledochocyst resection. The patient experienced repeat cholangitis with left hepatolithiasis during follow-up. Robotic left hepatectomy and revision of hepaticojejunostomy were performed smoothly. The patient recovered uneventfully and remained symptoms-free at a follow-up of 20 months. The robotic approach is beneficial in the fine dissection of the hepatic hilum and revision of hepaticojejunostomy in this particular patient

Robotic left hepatectomy with revision of hepaticojejunostomy

Laparoscopic hepatectomy and hepaticojejunostomy remain a surgical challenge despite the recent advances in minimally invasive surgery. A robotic surgical system has been developed to overcome the inherent limitations of the traditional laparoscopic approach. However, techniques of robotic hepatectomy have not been well described, and a description of robotic major hepatectomy with bilioenteric anastomosis can be found only in two previous reports. Here, we report a 33-year-old man with a history of choledochocyst resection. The patient experienced repeat cholangitis with left hepatolithiasis during follow-up. Robotic left hepatectomy and revision of hepaticojejunostomy were performed smoothly. The patient recovered uneventfully and remained symptoms-free at a follow-up of 20 months. The robotic approach is beneficial in the fine dissection of the hepatic hilum and revision of hepaticojejunostomy in this particular patient

Differential Transcriptomic Response in the Spleen and Head Kidney Following Vaccination and Infection of Asian Seabass with <i>Streptococcus iniae</i>

<div><p>Vaccination is an important strategy in the protection of aquaculture species from major diseases. However, we still do not have a good understanding of the mechanisms underlying vaccine-induced disease resistance. This is further complicated by the presence of several lymphoid organs that play different roles when mounting an immune response. In this study, we attempt to elucidate some of these mechanisms using a microarray-based approach. Asian seabass (<i>Lates calcarifer</i>) were vaccinated against <i>Streptococcus iniae</i> and the transcriptomic changes within the spleen and head kidney at one and seven days post-vaccination were profiled. We subsequently challenged the seabass at three weeks post-vaccination with live <i>S. iniae</i> and similarly profiled the transcriptomes of the two organs after the challenge. We found that vaccination induced an early, but transient transcriptomic change in the spleens and a delayed response in the head kidneys, which became more similar to one another compared to un-vaccinated ones. When challenged with the pathogen, the spleen, but not the head kidneys, responded transcriptomically at 25–29 hours post-challenge. A unique set of genes, in particular those involved in the activation of NF-κB signaling, was up-regulated in the vaccinated spleens upon pathogen challenge but not in the un-vaccinated spleens. A semi-quantitative PCR detection of <i>S. iniae</i> using metagenomic DNA extracted from the water containing the seabass also revealed that vaccination resulted in reduction of pathogen shedding. This result indicated that vaccination not only led to a successful immune defense against the infection, but also reduced the chances for horizontal transmission of the pathogen. In conclusion, we have provided a transcriptomic analysis of how the teleost spleen and head kidneys responded to vaccination and subsequent infection. The different responses from the two organs are suggestive of their unique roles in establishing a vaccine-induced disease resistance.</p></div

Establishing environmental DNA and RNA protocols for the simultaneous detection of fish viruses from seawater

Abstract Aquatic viruses are major threats to global aquacultural productivity. While conventional diagnostic methods for disease investigation are laborious, time‐consuming, and often involve the sacrifice of animals, environmental DNA and RNA (eDNA/eRNA) tools have the potential in being non‐invasive alternatives for the effective and early detection of various pathogens simultaneously. In this study, three seawater filtration methods—Sterivex syringe filtration, centrifugal ultrafiltration, and vacuum pump filtration with iron flocculation—were assessed for the recovery rates in co‐detecting fish virus eDNA/eRNA from natural seawater that was spiked with fish red seabream iridovirus (RSIV, DNA virus) and nervous necrosis virus (NNV, RNA virus). The centrifugal ultrafiltration method was the most effective for the capture of small‐sized viruses like NNV with a recovery rate of 63.23%, while the method of vacuum pump filtration with iron flocculation and chloroform disintegration of filter membranes had the highest RSIV recovery rate of 32.61%. We also optimized both automated and manual nucleic acid extraction methods and found comparable eDNA/eRNA extraction efficiencies. Our findings from the systematic comparison of seawater filtration and extraction methods suggest that each seawater filtration/nucleic acid extraction method can cater to different aquatic animal virus surveillance and disease investigation scenarios. These highlight the potential of virus eDNA/eRNA approaches for advancing the field of disease ecology and safeguarding aquatic animal health

Replication of SDDV on SK21 cells.

<p>SK21 cells were inoculated with an MOI of 0.01 TCID₅₀/cell SDDV (passage 3). (A/C) Negative control confluent monolayer of SK21 cells on day 3 (A) and 4 (C); (B/D) SDDV-infected SK21 cells on day 3 (B) and 4 (D) after inoculation; (E) SDDV genome copy number/mL (red line and left y-axis) and infectious titer (blue diamonds and right y-axis) of the infected SK21 cells in time. Error bars represent the standard deviation.</p

Protection of scale drop syndrome by vaccination of <i>Lates calcarifer</i>.

<p>Kaplan-Meier survival curve of fish vaccinated with formalin-inactivated virus (FK-SDDV), binary ethyleneimine-inactivated virus (BEI-SDDV), recombinant MCP protein (recMCP), or diluent control vaccine (placebo). Twenty-eight days after vaccination the fish were challenged with 2 x 10⁷ TCID₅₀ SDDV per fish (intraperitoneal), mortality was scored until day 28 after challenge. All three vaccines provided >70% RPS against disease (p<0.001, Tarone-Ware test).</p

Inoculation of SDDV in <i>Lates calcarifer</i>.

<p>(A) Kaplan-Meier survival curve of infected fish with 5x10⁶ TCID₅₀/fish (IP—black line) or 5.5 x10⁵ TCID₅₀/fish (IM—blue line) or both (IP + IM—yellow line). IP (1:10; green line) dose equaled 5.5 x10⁵ TCID₅₀/fish. (B) Control fish at day 10; (C) SDDV-infected fish (IP-high dose) at day 10 after infection. Note the fin erosion, tail erosion, body color variations, white or less mucus (inset), and changes in eye color. (D) SDDV genome copy number in serum of the fish on day 1, 7, 10, and 14 after infection. Diamond—black line: IP (mortality 60%); Triangle—blue line: IM (mortality 13%); Single cross—yellow line: IP + IM (mortality 47%); Square—green line: IP 1:10 (mortality 20%).</p

Vaccines.

<p>^a FK-SDDV: formalin-inactivated virus, BEI-SDDV: binary ethyleneimine-inactivated virus, recMCP-SDDV: recombinant MCP protein.</p><p>^b 0.1 mL vaccine administered per fish (IP), concentration is prior to inactivation for inactivated vaccines. N.A.: Not applicable.</p><p>Vaccines.</p