Abstract

The objective of this study was to determine the prevalence of Encephalitozoon cuniculi infection through serologic and histopathologic examination of laboratory rabbits from suppliers in India. One hundred and thirty-one New Zealand White rabbits procured from two different Indian suppliers were used for this study. All rabbits were clinically normal at receipt and during the period of experiment. Serological examination was carried out for the detection of E. cuniculi antibodies using ELISA tests. The select tissues were collected and processed for light microscopic evaluation. Among 131 rabbits evaluated 43 (33%) were seropositive for E. cuniculi. The microscopic changes in the brain and spinal cord (granulomatous inflammation and/or perivascular infiltrates) and kidneys (interstitial nephritis) were consistent with E. cuniculi infection. A good correlation (84%) between serological and microscopic findings was observed. Other background microscopic findings were minimal and consistent with hepatic (3%) and intestinal (5%) Eimeriasis (Coccidiosis). The microscopic changes consistent with otitis externa and otitis media, possibly related to external parasitic infection were observed in a few (11%) rabbits. Based on these findings, it is recommended that, rabbits should be serologically screened for E. cuniculi at suppliers breeding colony to remove carriers. Furthermore, before supplying rabbits to research facility, serology should be performed by the supplier to exclude seropositive animals. In addition, the research facilities also should consider performing serological testing to ensure that only seronegative animals are selected for experiments. This will minimize the variability in test results, avoid spurious observations and aid in scientific data interpretation

Abstract

Post approval monitoring (PAM) is an internal process for the ongoing research activities of approved protocols adopted by the institution. The primary goals are to ensure research methodology is conducted in accordance with the approval, evaluation of animal manipulations to reduce pain and distress, assessment of surgical procedures and post-operative care, to provide timely feedback to refine procedures and understand the proficiency of individuals on specific techniques of the species involved in experiments. The best approach should be collegial and facilitative of informed observations that provide an oversight of experiments conducted by principal investigators (PIs) along with other researchers listed in the protocol. In general, institutions can efficiently monitor this PAM process through any researcher/veterinarian/ coordinator who have considerable research experience as well as involved in the protocol review process or may be part of ethical committee for effective oversight of the animal care program. The proactive approach is fostering scientific conversation with research personnel and builds relationship by adhering institutional policies on animal research which in turn creates a positive culture of compliance throughout the institution. Collectively, PAM observation is ensuring the quality, research integrity, compliance with regulations and well-being of animals that eventually provides an opportunity for identifying educational competency, training based on observation categories and enabling the systematic communication process between animal care technicians, veterinarians, investigators, ethics committee and management for better compliance and humane care and use of animals for research.

Abstract

In the recent years, zebrafish (Danio rerio) species has been popularized as an animal model of human biology in understanding the genetic mechanism of vertebrate development and disease. Aquarium water is an immediate surrounding medium for the fish species in laboratory setup that determine their survival and normal physiological activities. Raising healthy zebrafish in the laboratory environment is prerequisite to ensure disease free stock for better research outcome. Important water parameters such as temperature, pH, electric conductivity, salinity, dissolved oxygen and CO₂ are required to be monitored regularly as a part of quality control. An episode of acute mortality was witnessed in several lines of laboratory zebrafish maintained at the facility of TIFR, Mumbai. The clinical symptoms include lethargy, swimming inability, settling down at the bottom and acute death. The acute mortality was evidenced irrespective of age, sex and strains of the zebrafish. The investigation was carried out to find the etiological agents implicated in the acute mortality. After ruling out various causative agents associated with the mortality, an investigation was zeroed on the pH of the fish aquarium water, the culprit for the acute death episode. The highly acidic pH (3.5 to 4.0) of the fish aquarium water was brought to normal level by adding buffer solution to avoid further mortality. Present paper describes an episode of acute death, timely diagnosis and successful measures taken to rescue the Danio rerio loss due to highly acidic aquarium water.

Abstract

Refinement of procedure plays a pivotal role on improving the well-being of laboratory animals that enables successful outcomes of experiments by obtaining reliable data. In general, identical practices reduce variability among mice and there may be several factors that can confound results which becomes inevitable within the context of long-term experiments. We explored several refinements pertaining to surgical techniques and husbandry practices of ongoing studies with the objectives of reducing early mortality during post-operative period (72h) and ensuring animal welfare as higher priority by providing better alternative husbandry practices until the termination of experiments. The entire refinement process was systematically implemented in mice where surgical intervention of bile duct ligation (BDL) was carried out and allowed to develop liver fibrosis as part of therapeutic requirement then treated with new chemical entities (NCEs) along with standard of care (SOC) upto 14 days. Survival surgery involving large number of mice in different groups was approved by the ethical committee. The notable refinements implemented such as suture patterns along with tissue adhesive to prevent wound dehiscence; replacement of corncob by adopting paper based bedding with nesting pads as enrichments; provision of Individually Ventilated Cages (IVC) with top inlet air supply for uniform flow in order to minimize hypothermia. Other contributing factors which included the use of improved surgical platform with thermo-controlled integral monitoring systems; Provision of infra-red light source immediately after surgery and until recovery; use of food supplement (Diet Gel) along with regular rodent diet; clinical scoring of mice during post-operative care and its management. However, there was no interference on fibrotic disease progression as well as experimental results because these refinements were implemented in a coordinated manner as part of ongoing process. Collectively, refinements of surgical as well as husbandry practices have improved the likelihood of mice and reduced incidences of mortality from 35% to 4% of four subsequent studies conducted at different intervals in mice.

Abstract

Abstracts presented during AFLAS 2018, Bangalore, India