Development of a Universal Scale for Validating the Process of Sanitization of Equipment, Materials and Surfaces in the Animal Facility

Background: Monitoring of sanitization of cage equipment is an essential function of any laboratory animal facilities, seeking to ensure the animal health and welfare. The purpose of the current study was to develop universal scale for monitoring sanitization through detection of residual ATP for the most effective process of sanitizing components of rodents micro- and macro-environment. Methods: Sterile pens and swabs for the HY-LiTE® Luminometer Instrument were used to measure ATP concentration (RLU) on the cleaned surface samples. We have examined the elements of the microenvironment (rodent and rabbit cage, cage wire meshes, feed separating pieces, water bottles with tips, houses, tunnels, IVC cage frames, cage tops and cage wire meshes). The assessed swab area on each of the surfaces was 10×10 cm. Result: We observed, that combined washing (cleaned manually and with automatic universal washers with detergent) gives the lowest RLU values. Monitoring of the quality of sanitization of equipment and surfacescan be carried out without the use of microbiological tests. Use of pre-washing allows increasing the service life of caging equipment. We have developed an assessment scale to monitor sanitizing, which can be recommended to scientific and breeding animal facilities for monitoring sanitization cage equipments.

Dexmedetomidine Associated Lung Cancer Proliferation and Tumor Growth is Prevented by β-Caryophyllene: Role of AMPK Activation and PGC-1α/TFAM Over Expression

Background: Dexmedetomidine has been reported to induce anti-apoptotic effects and metastatic progression in lung cancer. In the current investigation, the effect of β-Caryophyllene on dexmedetomidine induced cell proliferation and apoptosis of lung cancer cells and tumor growth in mice was studied. Methods: A549 cell line was cultured with either dexmedetomidine alone or together with β-Caryophyllene for 24 h and analysed for cell proliferation with MTT assay. ELISA based kit was used to determine apoptotic DNA fragmentation. Western blotting was used to determine expression levels of target proteins. The induction of experimental lung tumor in rat model was achieved through the injection of A549 tumor cells subcutaneously into the middle left side of the mice after anesthetization with pentobarbital (35 mg/kg) at 2.8 × 106 cells in 400 μl of PBS. Result: We found that β-Caryophyllene exerts the anti-proliferative effects on A549 cells. Furthermore, β-Caryophyllene significantly prevents apoptotic cell death and causes up-regulation of PGC-1α and TFAM compared to dexmedetomidine treated cells. We observed that β-Caryophyllene suppressed tumor development in mice significantly compared to dexmedetomidine treated group without changing body weight.

Anthelmintic Activity of Nine Varieties of Cajanus cajan (L.) Millsp on Haemonchus contortus from Sheep

Background: Internal parasitosis is the most important parasitism in small ruminants in tropical region. Synthetic anthelmintics are usually used for their control. Due to the emergence of resistance in worm population, the use of alternative methods such as plants bioactive molecules are developed. This study aimed at assessing anthelmintic activity of nine varieties of Cajanus cajan, a taniferous plant cultivated in West Africa. Methods: Leaves of the nine varieties of Cajanus cajan were dried, ground and hydroacetonic extracts were obtained by cold maceration at a concentration of 60 mg/ml. Live adult of Haemonchus contortus were obtained from sheep’s stomach and exposed, in triplicate, to the following solutions: hydroacetonic extracts (60 mg/ml), hydroacetonic extracts (60 mg/ml) associated with polyvinylpyrrolidone (PVP) (50 mg/ml), Levamisole (20 mg/ml) and phosphate buffered saline (PBS) solution. Worm death time was recorded for each treatment. Result: Worms exposed to Levamisole recorded the shortest death time about 9.73±1.77 min. The potent of the extracts induced worm death time of 64.83±4.73 min while the least efficient induced a death time of 156.50±23.20 min. Worms in the PBS solution were still alive after 24 hours of exposure. These results indicated that the four varieties of C. cajan used in this study have promising wormicidal. Besides that, the effect of tannins were not the only compound responsible for the anthelmintic activity.

Potential of Sambiloto (Andrographis paniculata Nees) Extract against Salmonella Pullorum

Background: Salmonella Pullorum are pathogenic bacteria that causes salmonellosis and causes heavy economic losses in the poultry industry and are zoonotic. Treatment of diseases caused by bacteria generally use antibiotics, but excessive administration of antibiotics causes bacterial resistance and residues in livestock. Major chemical constituents of Sambiloto are andrographolide and flavonoids. Andrographolide has antibacterial effect in addition to being antitoxic, anticancer, anti-inflammatory and antiallergic. Methods: The research was conducted by isolating and identifying Salmonella Pullorum on SSA media and a series of biochemical tests (TSIA, SIM, SCA, urea media and sugar test), manufacturing sambiloto extract, testing the sensitivity of several antibiotics using the disk diffusion method and testing the activation of sambiloto extract against Salmonella Pullorum using the disk diffusion and dilution methods. Result: The result show that sambiloto had antibacterial activity because it contained andrographolide, flavonoids, saponins, alkaloids and tannins and the lowest extract dose that effectively killed Salmonella Pullorum is concentrations of 20%.

Effects of Water Temperature on Activities of Metabolic Enzymes of Juvenile Siganus guttatus

Background: Temperature is one of the most important environmental factors affecting the survival, growth and metabolism of fish. The current study was aimed to study the effects of water temperature on the metabolic enzyme activities of juvenile Siganus guttatus. Methods: The juveniles were domesticated at 28±1°C for two weeks and then the temperature was adjusted to the target temperature groups (31°C, 27°C, 23°C and 19°C) by the gradually increasing or decreasing temperature with the change rate of 2°C per day. The experiment lasted for 70 d. At the end of the experiment, the fish were anesthetized and all the livers were dissected on ice plate and stored in the refrigerator at -80°C for the determination of enzyme activity. Result: The activities of glutamic pyruvic transaminase (GPT) and glutamic oxaloacetic transaminase (GOT), hexokinase (HK) and pyruvate kinase (PK), lipoprotein lipase (LPL) and hepatic lipase (HL) tend to be increased with the reduction of temperature. The above enzymes activities in 19°C group were highest. The activity of lactate dehydrogenase (LDH), succinate dehydrogenase (SDH) and citrate synthase (CS) was lowest in 19°C. These results suggests that 19°C had exceeded the suitable temperature range for juvenile S. guttatus. At low temperature, S. guttatus mainly use fat for energy, but less anaerobic metabolism for energy.

Detection of a New Variant in Outbreak of Bovine Ephemeral Fever in Turkey, 2020

Background: In this study, partial nucleotide sequence analysis of the G gene was performed for the molecular characterization of the virus that caused the bovine ephemeral fever virus (BEFV) epidemic in Turkey in 2020. Phylogenetic analysis of these nucleotide sequences was performed with the virus nucleotide sequences of the epidemics seen in 2008 and 2012. These sequences were announced in GenBank. Phylogenetic analysis of these nucleotide sequences was performed with the virus nucleotide sequences of the epidemics seen in 2008 and 2012. Methods: The study was conducted in dairy cattle holdings located in Diyarbakır Sur, Çınar and Dicle regions in South-eastern Turkey in August-November 2020. The number of animals in the holdings consisted of 750 (n=750), 150 (n=150) and 200 (n=200) cattle, respectively. Result: Severe respiratory symptoms and high mortality in the affected animals were notable symptoms. As a result of the phylogenetic analysis, it was determined that the virus that caused the epidemic in Turkey in 2020 was formed by a new variant in the Turkey-2 group, which was similar to the Indian isolates, unlike the Turkey-1 group, which was close to the Middle East variants in 2008 and 2012 isolates.

Comparison of Different Count Models for Investigation of Some Environmental Factors Affecting Stillbirth in Holsteins

Background: The objective of this study is comparing different count data models for stillbirth data. In modeling this type of data, Poisson regression or alternative models can be preferred. Methods: The poisson, negative binomial, zero-inflated poisson, zero-inflated negative binomial, poisson-logit hurdle and negative binomial-logit hurdle regressions were compared and used to examine the effects of the gender, parity and herd-year-season independent variables on stillbirth. Furthermore, the Log-Likelihood statistics, Akaike Information Criteria, Bayesian Information Criteria and rootogram graphs were used as comparison criteria for performance of the models. According to these criteria, Negative Binomial-Logit Hurdle Regression model was chosen as the best model. Result: The parameter estimates obtained by Negative Binomial-Logit Hurdle Regression model in relation to the effects of the gender, parity and herd-year-season independent variables on stillbirth were found to be significant (p less than 0.01). It was found that while stillbirth incidence was higher in males than females, it was found to decrease as the parity increased. As a result, the Negative Binomial Logit Hurdle model was found the best model for stillbirth count data with overdispersion.

Effect of Body Condition Score at Calving on Postpartum Performance of Murrah Buffaloes

Background: Body condition score (BCS) system is a subjective method to assess the body fat reserves particularly over the bony prominences like back and pelvic region. It is based on evaluation of the outer appearance of the animal that interacts with its body fat reserves and therefore is directly influenced by energy balance. Studies on the relationship between BCS and postpartum performance were meager in buffaloes. Hence, the present work was carried out to study the relationship between BCS at calving and postpartum performance in Murrah buffaloes. Methods: The effect of body condition score at calving (BCSc) on the postpartum performance was studied in 24 Murrah buffaloes grouped based on BCS, maintained at Buffalo Research Station during 2018-19. Result: The study showed that buffaloes of BCSc above 3.5 with significantly (P less than 0.01) higher body weights showed better reproductive performance compared to buffaloes of BCSc groups below 3.5 with less body weights. The prepartum and weekly postpartum changes in BCS studied showed that the buffaloes gained a BCS of 0.57 from 3 months prepartum to one week prepartum, lost 0.17 BCS due to calving, further showed a decline in BCS up to 9 weeks of postpartum and then started regaining BCS gradually until 18 weeks postpartum. The mean LBCS over the period of 18 weeks postpartum was comparatively higher (0.75±0.05) than BCS restored in the early lactation which was shown as GBCS (0.34±0.02). Buffaloes of BCSc group 3.5-3.99 showed the best reproduction performance among the four BCSc groups with less postpartum estrus period (46.66 days), service period (58.83 days), number of service per conception (1.50) and high rate of 1st service conception (66.66%).

Evaluation of Ketamine Hydrochloride in Combination with Midazolam, Dexmedetomidine and Butorphanol as Balanced Anaesthesia in Cats

Background: As cats undergo stress at the time of presentation before surgery and therefore sedation is required often. It is very difficult to choose a balanced anaesthetic protocol during the surgical intervention in the case of a cat. Therefore, the present study was undertaken to evaluate the effect of ketamine hydrochloride in combination with midazolam, dexmedetomidine and butorphanol as balanced anaesthesia on clinico-physiological, haematological, serum biochemical profile in the surgical management of cats undergoing neutering. Methods: The study was performed on 18 clinical cases of cats which were brought to the Teaching Veterinary Clinical Complex, College of Veterinary Sciences and Animal Husbandry (CVSc and AH), Central Agricultural University (CAU), Selesih, Aizawl, Mizoram for elective surgery such as neutering. Cats were randomly divided into 3 equal groups, i.e. Gr. A, Gr. B and Gr. C. Animals were premedicated with glycopyrrolate @ 0.01 mg/kg, i/m. After 10 minutes of premedication, in Gr. A ketamine hydrochloride @15 mg/kg and midazolam @ 0.5 mg/kg, in Gr. B ketamine hydrochloride @10 mg/ kg and dexmedetomidine@ 5 mcg /kg and in Gr. C ketamine hydrochloride @ 5 mg/kg, dexmedetomidine@ 5 mcg/kg and butorphanol @ 0.2 mg/kg was administered intramuscularly. Clinico-physiological and haemato-biochemical profiles were evaluated at 0 minute (baseline), then at 15, 30 and 60 minutes after administration of anaesthetic agents to evaluate their anaesthetic effect. Result: In the case of time for induction, the quality of induction, assessment of peri-operative analgesia, depth of anaesthesia and quality of recovery there was no significant difference among all three groups. In Gr. B, animals showed significantly higher duration of recumbency (DOE) and recovery. The assessment of peri-operative analgesic effect among the groups revealed that analgesia during the perioperative period appeared best in Gr. C protocol. In regards to depth of anaesthesia, ketamine hydrochloride in combination with dexmedetomidine and butorphanol group showed a better result. Rectal temperature decreased significantly (P greater than 0.05) up to 60th minutes in Gr. A, but it remained within the physiological range. Heart rate raised significantly (p less than 0.05) from the baseline (0 minutes) onwards and decreased significantly (p less than 0.05) till the end (60 minutes) of the study in Gr. C. The respiration rate was significantly (p less than 0.01) different in between time intervals for all three groups. No significant difference was observed on haemato-biochemical observation except glucose level which significantly increased after induction of anaesthesia. In conclusion, treatment with ketamine hydrochloride in combination with dexmedetomidine and butorphanol led to acceptable sedation and mild changes in clinico-physiological, haematological, serum biochemical profile in comparison to the other two groups.

​​Nursery Rearing of Penaeus vannamei in Biofloc Systems with Different Salinities and Organic Carbon Sources

Background: Nursery rearing of Penaeus vannamei became inevitable in the Indian shrimp farming industry, since intensification of culture practices in grow-out systems caused nitrogenous wastes accumulation, diseases, mortality and premature harvests resulted in high food conversion ratio, lower production and profits. The nursery rearing in traditional water exchange systems often getting failure because of nitrogenous waste accumulation. Hence, the present experiment was planned to rear P. vannamei in bioflocs systems (BFS) with different salinities and carbon sources. Methods: The experiment was conducted in 0.22 m3 tanks with three different salinity groups viz., 35 ppt, 20 ppt and 5 ppt and in each salinity sugar, molasses used as carbon sources to maintain an estimated C/N ratio of 15:1 and controls without carbon sources. Experimental tanks were stocked @6 post larvae/l, with pre-salinity acclimatized P.vannamei seeds having 0.0029±0.0003g size and nursed for a period of 5 weeks. Result: The nitrogenous waste accumulation was reduced significantly (p less than 0.05), also average body weight and survival rate of the seeds showed significant difference (p less than 0.001) between treatments and controls and within treatments (p less than 0.05). Salinity, carbon sources and their interaction influenced the growth characteristics significantly (p less than 0.01). The present experiment manifested promising results of bioflocs nurseries in rearing of P. vannamei seeds at different salinities.